\ 


THE  VITAMINES 


BY 


CASIMIR  FUNK 

** 
ASSOCIATE  IN  BIOLOGICAL  CHEMISTRY 

College  of  Physicians  and  Surgeons 
New  York  City 


Authorized  Translation 

from 
Second  German  Edition 

BY 

HARRY  E.  DTJBIN 

FORMERLY  ASSISTANT  IN  RESEARCH  MEDICINE 
University  of  Pennsylvania 
Philadelphia,  Pa. 


BALTIMORE 

WILLIAMS  &  WILKINS  COMPANY 
1922 


COPYRIGHT  1922 
WILLIAMS  &  WILKINS  COMPANY 

Made  in  United  States  of  America 

The  right  of  publication  in  English 
is  reserved 


32NTAL  DEPARTMENT 


COMPOSED   AND  PRINTED  AT  THE 

WAVERLY  PRESS 

BY  THE  WILLIAMS  &  WILKINS  COMPANY 

BALTIMORE,  MD.,  U.  S.  A. 


CONTENTS 

Preface  to  the  first  edition 9 

Preface  to  the  second  edition 11 

Translator's  note 13 

Introduction 15 

Historical  survey 19 

Introductory 35 

PART  I 
THE  VITAMINE  REQUIREMENTS  OF  PLANTS  AND  ANIMALS 

Chapter  I.     The  role  of  the  vitamines  in  the  vegetable  kingdom 49 

Yeast 51 

Bacteria 56 

Fungi 64 

Higher  plants 65 

Chapter  II.  The  role  of  the  vitamines  in  the  animal  kingdom 73 

Life  without  bacteria 76 

Protozoa 79 

Metazoa 81 

Growth  of  tissue  in  vitro 81 

Insects 83 

Fish 85 

Amphibia 85 

Birds 87 

Chickens — normal  nutrition ;  chicken  beriberi 87 

Pigeons — beriberi 95 

Pathological  anatomy  and  chemical  pathology  of  beriberi  in  birds.  103 

Mammals 109 

Rats 110 

Mice 125 

Guinea  pigs 127 

Rabbits 134 

Cats 135 

Lions 136 

Dogs 136 

Domestic  animals — sheep,   goats,   reindeer,   pigs,   horses,   cows, 

oxen,  (stijfziekte,  lamziekte,  pica) 140 

Monkeys 153 

Man..                                                                                                       .  155 


4  CONTENTS 

PART  II 
CHEMISTKY,  PHYSIOLOGY  AND  PHAEMACOLOGY  OF  THE  VITAMINES 

Chapter  I.  The  antiberiberi  vitamine — Vitamine  B 163 

Chemical  investigation  of  rice  polishings 167 

Chemical  investigation  of  yeast 177 

Other  sources  of  supply 192 

Synthetic  experiments  — 193 

Stability  of  the  antiberiberi  vitamine  against  heat  and  chemical  and 

physical  agents 194 

Demonstration  and  estimation  of  antiberiberi  vitamine 195 

The  possible  identity  of  vitamine  B  with  the  substance  stimulating 

the  growth  of  animals  and  of  yeast 203 

Specificity  of  antiberiberi  vitamine 205 

Physiology  and  pharmacology  of  antiberiberi  vitamine 207 

The  influence  of  the  dietary  composition  on  the  vitamine  requirement .  210 

Chapter  II.  The  antirachitic  vitamine — Vitamine  A 218 

Chemistry  of  cod  liver  oil 218 

Chemistry  of  vitamine  A  in  butter  and  other  sources 220 

Nature  of  vitamine  A 222 

Demonstration  of  vitamine  A 225 

Chapter  III.  Relationship  of  the  antiberiberi  and  antirachitic  vitamines 

to  lipoids 228 

Chapter  IV.  The  antiscorbutic  vitamine — Vitamine  C 231 

The  chemistry  and  the  nature  of  vitamine  C 231 

Chapter  V.  Vitamine  content  of  various  foodstuffs  in  the  natural  and 

prepared  condition 236 

Influence  of  heating  and  cooking  on  the  vitamine  content 238 

Vitamine  content  of  the  cooking  water 241 

Influence  of  drying 241 

Influence  of  canning 242 

Influence  of  ageing  and  storing 243 

Cereals 244 

Rice 248 

Barley 248 

Oats 249 

Wheat 249 

Maize 250 

Potatoes 255 

Milk 256 

Demonstration  of  vitamines  in  milk 256 

Influence  of  food  on  the  vitamine  content  and  composition  of  milk  257 

Vitamine  content  of  milk 258 

Influence  of  heat  on  the  vitamine  content  of  milk 259 

Influence  of  drying  and  evaporation 260 

Nutritive  value  of  milk. .  261 


CONTENTS  5 

Meat 262 

Table  showing  the  vitamine  content  of  the  most  frequently  used 
foodstuffs 263 

PART  III 

THE  HUMAN  AVITAMINOSES — CONDITIONS  IN  WHICH  THE  VITAMINES 

PLAY  A  R6LE 

Chapter  I.  Beriberi 275 

Mode  of  occurrence  of  and  diets  leading  to  beriberi 280 

Symptomatology  and  types  of  beriberi 282 

The  sensory,  motor  form 283 

The  dry,  atrophic  form ' 287 

The  wet,  atrophic  form. 287 

Epidemic  dropsy 288 

The  pernicious,  acute  form 289 

Infantile  beriberi 290 

Pathology 291 

General 291 

.  Blood «.  293 

Cerebrospinal  fluid 293 

Urine 293 

Therapy 293 

General 293 

Infantile 294 

Ship  beriberi 294 

Relationship  between  beriberi  and  scurvy 295 

Chapter  II.  Scurvy 297 

Mode  of  development 300 

Symptomatology  and  progress  of  scurvy 303 

Infantile  scurvy 306 

Diagnosis 307 

Hematology 308 

Metabolism  experiments 308 

Pathology 308 

Barlow's  disease 309 

Therapy 311 

Chapter  III.  Rickets 312 

Occurrence 313 

Symptomatology  and  diagnosis 315 

Pathological  anatomy  and  chemical  pathology  of  rickets 315 

Metabolism 317 

Therapy  and  therapeutic  effect  on  metabolism 220 

Etiology 323 

Vitamine  etiology  of  rickets 324 


6  CONTENTS 

Osteomalacia 329 

Symptoms 330 

Anatomy 330 

Metabolism 331 

Therapy 331 

Etiology 331 

Chapter  IV.  Some  nutritional  disturbances  in  children — tetany,  carbohy- 
drate dystrophy  (Mehlnahrschaden),  atrophy 332 

Tetany  (Spasmophilia) 332 

Etiology 332 

Symptoms 333 

Therapy 334 

Carbohydrate  dystrophy  (Mehlnahrschaden) 335 

Etiology 336 

Atrophy 336 

Other  nutritive  disturbances  of  children 337 

Chapter  V.  Nutrition  in  man — An  introduction  to  the  study  of  pellagra 

and  hunger  edema 340 

Chapter  VI.  Pellagra 351 

Geographical  distribution 351 

Progress  of  pellagra ". . .  352 

Acute,  malignant  form 354 

Light,  sub-chronic  form 354 

Severe,  cachetic  sub-chronic  form 354 

Chronic  form 354 

Symptomatology  and  pathology 355 

Gastro-intestinal  tract 355 

Skin 357 

Nervous  system 357 

Circulatory  system 359 

Bones 360 

Sexual  organs 360 

Other  organs 361 

Chemical  pathology 361 

Metabolism ...... 362 

Prognosis 362 

The  relationship  of  pellagra  to  the  accepted  avitaminoses — beriberi 

and  scurvy 362 

Mode  of  development  of  pellagra 363 

Therapy 366 

Etiology 366 

Chapter  VII.  Sprue 369 

Symptomatology 369 

Pathological  anatomy 370 

Therapy 370 

Pathogenesis 371 


CONTENTS  7 

Chapter  VIII.  Hunger  edema 372 

Symptomatology  and  mode  of  development 373 

Pathology 374 

Metabolism 375 

Therapy S75 

Etiology 376 

Chapter  IX.  Pathological  conditions  in  which  the  lack  of  vitamines  may 

be  suspected 378 

Kallak 378 

Trench  Sickness 378 

Intestinal  stasis,  etc 379 

Sterility 379 

Hemeralopia 379 

Exophthalmic  goitre  (Graves'  Disease), 380 

Significance  of  vitamines  in  infections 380 

Ophthalmia 381 

Nature  of  the  disease 382 

Tuberculosis 384 

Leprosy 385 

Pneumonia 385 

Chapter  X.  Influence  of  nutrition   (vitamines)   on  the  action  of  some 

poisons  and  upon  pathological  conditions  of  non-infectious  origin  . . .  386 

Anemia,  etc  386 

Diabetes 387 

Cancer 388 

Development  of  teeth 391 

Calculi 393 

General  literature  on  the  subject  of  the  vitamines 395 

Literature  to  the  text 399 

Index  to  Sectional  Divisions  of  the  Literature 476 

Author  index 477 

Subject  index 491 


PREFACE  TO  THE  FIRST  EDITION 

This  initial  attempt  at  a  classification  of  our  knowledge  on  vita- 
mines  and  avitaminoses  contains,  besides  a  number  of  facts,  the  per- 
sonal conceptions  of  the  author,  new  questions,  ideas  and  lines  of 
research.  Much  of  the  material  is  fragmentary  while  some  of  it  is 
purely  hypothetical.  This  small  book  is  to  be  regarded  as  the  first 
step  in  a  new  direction  in  the  field  of  physiology  and  pathology. 
Many  facts  are  still  lacking  in  this  field  of  work  so  that  hypotheses 
must  naturally  fill  the  gaps.  Even  if  the  structure  is  skeleton-like, 
at  least  the  fundamentals  are  well  established. 

At  this  point,  I  want  to  thank  Dr.  F*aser,  Dr.  Stanton,  Prol 
Hoist,  Dr.  Mott,  Dr.  Roberts,  Dr.  Standwith  and  Dr.  Zeller,  and  the 
London  Society  of  Tropical  Medicine,  for  the  permission  to  reproduce 
illustrations.  I  am  thankful  also  to  Dr.  Macauley  of  Cape  Town  for 
having  called  my  attention  to  a  South  African  cattle  disease.  In 
particular,  do  I  wish  to  express  my  deepest  gratitude  to  my  Father, 
Dr.  J.  Funk,  for  his  unvaried  and  tireless  assistance. 

I  would  appreciate  it  greatly  if  the  investigators  who  are  conducting 
research  on  problems  discussed  in  this  book  would  be  kind  enough 
to  send  me  reprints  of  their  work  which,  at  times,  is  available  only 
with  great  difficulty. 

THE  AUTHOR, 

Cancer  Hospital  Research  Institute, 
Brompton,  London,  S.  W. 

October,  1913. 


PREFACE  TO  THE  SECOND  EDITION 

This  edition,  totally  revised  and  almost  completely  re-written, 
appears  seven  years  after  the  first  edition,  deferred  because  of  the 
World  War.  In  the  interim,  we  have  received  many  communications 
from  our  colleagues  informing  us  that  our  effort  had  provided  them 
with  a  welcome  stimulus  to  their  work.  We  have  watched  the  great 
development  of  this  field  of  research  since  the  appearance  of  The 
Vitamines  and  it  is  a  source  of  great  pleasure  that  our  work  has  also 
contributed  to  this  progress.  The  views  expressed  at  the  time  have 
been  tested,  for  the  most  part,  and  found  correct.  While  the  first 
edition  was  published  at  a  time  when  great  differences  of  opinion 
prevailed,  our  desire  then  being  to  take  cognizance  of  all  of  them,  we 
feel  justified  now  in  reviewing  only  those  investigations,  the  view- 
point of  which  is  not  too  far  removed  from  ours.  In  this  way,  it 
has  been  possible  to  make  the  book  more  comprehensive.  The 
subject  of  vitamines,  already  beyond  the  stage  of  hypothesis,  is  based 
on  a  firm  foundation  and  has  received  universal  recognition.  In 
spite  of  this,  however,  we  are  well  aware  that  great  gaps  exist  in  our 
knowledge,  so  that  we  can  not  regard  the  chapter  as  closed. 

These  gaps  may  be  explained  by  the  fact  that  the  field  of  vitamine 
research  gave  many  investigators  the  opportunity  of  making  a  name 
for  themselves  with  rather  superficial  work.  Only  seldom  did  they 
seriously  endeavor  to  get  at  the  basis  of  the  phenomena  observed. 
To  permit  of  progress  in  the  subject  of  vitamines,  it  would  be  very 
desirable  that  at  least  some  of  the  workers  should  abandon  the  beaten 
path  of  exclusive  animal  experiments  and  break  a  fresh  trail  leading 
towards  the  chemistry,  physiology  and  pharmacology  of  the  vitam- 
ines, as  well  as  the  anatomical  pathology  of  the  avitaminoses. 

For  help  in  the  preparation  of  this  edition,  we  are  indebted  to 
many  of  our  colleagues;  we  wish  also  to  express  our  thanks  to  the 
Medical  Research  Committee  (London)  and  to  Dr.  Alfred  F.  Hess 
(New  York)  for  permission  to  reproduce  a  number  of  illustrations. 
In  addition,  we  wish  to  thank  those  whose  permission  it  was  impos- 
sible to  obtain  because  of  uncontrollable  circumstances.  We  are 
especially  grateful  to  Dr.  Richard  Hamburger,  First  Assistant  at  the 
Pediatric  Clinic,  Berlin,  who  took  it  upon  himself  to  correct  and 
critically  review  the  proofs. 

11 


12  PREFACE   TO   SECOND   EDITION 

In  particular,  however,  we  take  pleasure  in  recording  our  apprecia- 
tion of  the  assistance  extended  by  the  Hon.  Herman  A.  Metz  (New 
York)  who,  although  a  layman  in  this  field  of  work,  understood  its 
significance  and  gave  us  the  opportunity,  after  an  enforced  interrup- 
tion of  many  years,  of  resuming  the  experimental  work  in  the 
"Research  Laboratory  of  H.  A.  Metz." 

This  book  will  be  of  interest  not  only  to  nutrition  investigators 
and  childrens'  specialists,  but  to  every  physician  who  comes  into 
touch  with  questions  of  the  physiology  of  nutrition.  It  treats  many 
problems  which  are  closely  allied  to  other  lines  of  research  and  will 
be  of  value  to  plant  physiologists,  bacteriologists  and  animal  breeders. 

CASIMIR  FUNK. 

College  of  Physicians  and  Surgeons, 

Columbia  University, 

New  York  City. 

December,  1921. 


TRANSLATOR'S  NOTE 

The  translator  has  been  associated  with  the  author  for  three  years 
in  experimental  vitamine  research,  and  it  is  a  pleasure  to  record  at 
this  point  the  spirit  which  animates  him  in  his  work  and  his  earnest 
desire  to  see  real  progress  made  towards  the  solution  of  the  many 
problems  of  nutrition.  It  was  this  which  prompted  the  preparation 
of  the  present  edition,  intended  rather  as  a  stimulus  to  thought  and 
further  research  than  as  an  elaboration  of  technicalities.  Inasmuch 
as  the  author  has  had  the  opportunity  of  reading  the  manuscript 
before  it  went  to  press,  the  reader  may  be  sure  that  the  sense  of  the 
book  has  been  accurately  preserved. 

In  conclusion,  the  undersigned  wishes  to  extend  his  thanks  to  the 
Hon.  Herman  A.  Metz  for  the  many  facilities  provided  in  the  prepara- 
tion of  this  book. 

HARRY  E.  DUBIN. 

Research  Laboratory  of  H.  A.  Metz, 
New  York  City. 

December,  1921. 


13 


INTRODUCTION 

The  first  edition  of  this  book,  drafted  seven  years  ago,  was  intended 
to  serve  the  purpose  of  directing  the  attention  of  the  medical  and 
chemical  professions  to  this  new  and  attractive  field  of  investigation. 
The  findings  upon  which  we  based  our  views  in  1913  appeared  indeed 
to  be  far  from  indisputable  but  time  has  shown  the  comparative 
accuracy  of  our  conception.  Since  then,  we  have  made  it  a  point 
to  note  any  work  that  might  have  some  bearing  upon  the  question 
of  the  vitamines,  and  have  now  undertaken  to  point  out  what  may 
be  regarded  as  erroneous  and  what  may  be  looked  upon  as  correct, 
in  the  light  of  the  present  status  of  the  subject. 

Our  original  classification  included  beriberi,  scurvy  and  Barlow's 
disease  as  true  avitaminoses,  while  pellagra,  sprue,  rickets,  and  some 
metabolic  diseases  of  animals,  were  held  to  be  hypothetical  avitam- 
inoses. At  present,  beriberi,  scurvy  and  Barlow's  disease  are  uni- 
versally accepted  as  true  avitaminoses,  while  rickets  is  quite  generally 
acknowledged  as  such.  The  etiology  of  pellagra  and  sprue,  on  the 
other  hand,  has  not  yet  been  definitely  established.  Regarding 
pellagra,  which  we  shall  discuss  later,  the  dietetic  hypothesis  is  at 
present  in  the  foreground. 

As  for  the  chemistry  of  the  vitamines,  an  examination  of  the 
literature  reveals  little  progress  in  this  direction.  To  be  sure,  one 
must  bear  in  mind  that  the  outbreak  of1  the  war  was  not  particularly 
conducive  to  the  long  and  tedious  research  that  is  necessary,  if  this 
phase  of  the  subject  is  to  be  cleared  up.  From  tune  to  time,  word 
comes  from  this  or  that  laboratory  to  the  effect  that  the  puzzle  has 
been  solved — that  progress  has  been  made  even  towards  the  synthesis 
of  the  vitamines — but  after  much  patient  waiting,  nothing  more  is 
heard  of  the  discovery.  However,  since  a  large  part  of  the  experi- 
mental work  is  being  carried  on  in  many  industrial  research  labora- 
tories, it  is  quite  possible  that  definite  important  progress  has  been 
made,  of  which  reports  have  not  yet  been  published. 

Incidentally,  the  war  has  given  added  impetus  to  vitamine  research 
in  another  direction.  The  vitamines  have  now  become  of  great 
importance  from  the  point  of  view  not  only  of  pure  science  and  medi- 
cine, but  also  of  political  economy — much  greater  than  could  have 

15 


16  THE   VITAMINES 

been  foreseen  when  the  first  edition  of  this  book  appeared.  The 
war,  first  of  all,  prevented  the  normal  interchange  of  foodstuffs 
among  the  various  countries.  Subsequently,  this  was  followed  by  a 
disturbance  in  the  entire  mode  of  living,  due  to  the  necessity  of 
making  war,  the  utilization  of  farm  labor  for  other  purposes,  and  the 
lack  of  means  of  transportation.  Although,  as  this  is  being  written, 
more  than  two  years  have  elapsed  since  the  end  of  the  war,  conditions 
have  become  worse  instead  of  better,  according  to  available  reports. 
Similarly,  notwithstanding  the  national  rationing  of  food  both 
during  and  after  the  war,  the  people  have  suffered  because  of  the 
mistakes  made  during  the  war.  In  the  face  of  higher  prices,  general 
unrest  and  unwillingness  to  settle  down  again  to  some  productive 
work,  the  abnormal  conditions  already  mentioned  will  likely  prevail 
for  several  years  to  come.  It  is  not  our  purpose  to  suggest  that 
knowledge  of  the  vitamines  will  solve  the  present  difficulties.  Still, 
we  shall  call  attention  to  facts  and  principles  which,  at  present,  are 
of  universal  interest — principles  which  may  contribute  to  the  allevia- 
tion of  the  wretchedness  of  stricken  Europe. 

Considering  more  closely  the  dietaries  of  farmers  and  of  the  rural 
populations  in  various  parts  of  the  world,  it  is  easy  to  see  that  no 
knowledge  of  vitamines  is  necessary  to  keep  those  people  in  good 
health.  From  generation  to  generation,  their  nutrition  has  been 
regulated  according  to  the  climate,  the  economic  situation,  and  the 
exigencies  of  the  work  performed.  Of  course,  in  some  oriental 
countries,  where  conditions  are  not  so  well  known  to  us,  instinct  some- 
times does  not  choose  the  correct  food  and  hence  the  prevalence  of 
beriberi.  All  in  all,  we  see  that  the  white  races  have  a  wholesome 
knowledge  of  their  food  requirements,  which  is  only  natural.  In 
certain  provinces,  before  the  war,  it  was  possible  to  see  examples  of 
particularly  monotonous  and  simple  dietaries  accompanied,  on  the 
whole,  by  no  pathological  conditions.  We  may  be  sure,  however, 
that  since  these  same  peoples  apparently  subsisted  on  a  practically 
unchanging  diet  for  hundreds  of  years,  they  would  most  certainly  not 
have  survived  if  their  choice  of  diet  had  been  anything  but  correct. 

When  the  usual  equilibrium  is  disturbed  by  extraneous  conditions 
such  as  war,  a  financial  crisis  or  a  catastrophe,  then  the  practical 
knowledge  of  centuries  suddenly  becomes  useless  to  the  people,  and 
they  are  obliged  to  seek  some  other  basis  of  existence.  Untold 
hardships  are  endured  till  this  is  accomplished,  the  population  func- 


INTRODUCTION  17 

tioning  similar  to  experimental  animals,  used  to  establish  the  value 
of  new  foodstuffs.  Here,  at  least,  there  is  the  added  advantage  of 
successfully  applying  the  various  principles  of  nutrition  and  sharply 
terminating  the  unnecessarily  protracted  period.  In  this  connection, 
it  is  not  impossible  to  make  immediate  use  of  the  enormous  amount 
of  information  obtained  on  the  subject,  both  before  and  during  the 
war,  especially  by  the  United  States  and  England.  During  the  past 
few  years,  virtually  all  foodstuffs  have  been  tested  as  to  their  nutritive 
value  and  their  vitamine  content.  Although  most  of  the  data  secured 
are  the  results  of  animal  experiments,  they  are,  with  a  few  reserva- 
tions, directly  applicable  to  man — especially  as  it  has  been  demon- 
strated that  there  are  no  vital  differences  between  the  findings 
obtained  with  man  and  those  with  animals. 

In  general,  it  may  be  said  that  during  the  war,  and  after,  there 
was  a  disarrangement  of  the  nutritional  elements  resulting  in  a 
decreased  consumption  of  protein  and  fat  and  a  greater  intake  of 
carbohydrate,  together  with  a  diminution  of  vitamines.  It  should 
be  noted  here  that  the  more  important  dietary  constituents,  in  com- 
parison with  the  physiologically  inferior  ones,  have  increased  greatly 
in  price,  so  that  there  is  this  danger  to  contend  with  in  addition  to 
the  element  of  scarcity.  Now,  when  a  well-planned  animal  experi- 
ment is  undertaken,  all  factors  are  controlled  as  far  as  possible  except 
that  one  whose  influence  upon  the  organism  is  being  determined. 
Quite  another  state  of  affairs  confronts  us  in  the  case  of  nutrition 
investigations  on  man,  where  conditions  arise  which  are  only  rarely 
met  with  in  animal  experiments.  If  there  is  a  lack  of  one  constituent, 
then  it  is  almost  certain  that  the  entire  choice  of  diet  is  not  correct. 
For  example,  if  the  protein  content  of  the  diet  is  too  low,  it  is  at  the 
same  time  apparent  that  the  carbohydrates  are  present  in  excess, 
that  the  vitamine  content  is  diminished,  and  that  the  inorganic 
elements  are  inadequately  grouped.  It  is  easily  possible  that  right 
at  this  point  is  where  we  must  search  for  an  explanation  of  the 
pathologically  occurring  avitaminoses,  the  etiology  of  which  it  is  so 
difficult  to  establish.  If  in  such  a  case  as  mentioned  above,  the 
missing  factor  in  the  diet  is  supplied,  good  results  do  not  follow,  for  the 
reason  that  still  other  factors  have  not  been  taken  into  consideration. 

Under  certain  circumstances,  a  seemingly  well  chosen  diet  may 
prove  to  be  inadequate,  particularly  when  special  demands  are  made 
upon  the  individual,  such  as  hard  work,  growth,  birth,  and  nursing. 


18  THE    VITAMINES 

All  these  factors  must  be  taken  into  account  when  commenting  upon 
the  pathological  conditions  which  are  of  exceptional  interest  to  us. 
These  are  chiefly  the  conditions  which  give  us  an  insight  into  the 
causes,  which  are  so  difficult  to  determine  in  hunger  edema  and 
pellagra.  Even  a  well  informed  physician  may  easily  be  led  astray. 
He  questions  his  patients  about  their  diet,  whereupon  they  enumerate 
a  long  list  of  foodstuffs,  from  which  apparently  nothing  has  been 
omitted.  Immediately,  his  attention  will  be  directed  to  obvious 
things,  such  as  mode  of  living,  nature  of  work  performed,  and  method 
of  preparing  and  cooking  the  food — everything  which  may  be  etio- 
logically  important  and  may  help  him  solve  his  problem. 

Despite  the  fact  that  a  number  of  ideas  originated  by  us  are 
credited  to  others,  it  is  a  source  of  pleasure  to  witness  the  great 
progress  that  has  been  made  in  vitamine  research.  In  our  opinion, 
the  name  "Vitamine",  proposed  by  us  in  1912,  contributed  in  no 
small  measure  to  the  dissemination  of  these  ideas.  The  word, 
"Vitamine",  served  as  a  catch  word  which  meant  something  even  to  the 
uninitiated,  and  it  was  not  by  mere  accident  that  just  at  that  time, 
research  developed  so  markedly  in  this  direction. 

Our  view  as  to  the  fortunate  choice  of  this  name  is  strengthened,  on 
the  one  hand,  because  it  has  become  popular  (and  a  badly  chosen 
catchword,  like  a  folksong  without  feeling,  can  never  become  pop- 
ular), and  on  the  other,  because  of  the  untiring  efforts  of  other  workers 
to  introduce  a  varied  nomenclature,  for  example,  "accessory  food 
factors,  food  hormones,  water-soluble  B  and  fat-soluble  A,  nutramine, 
and  auximone"  (for  plants).  Some  of  these  designations  are  cer- 
tainly not  better,  while  others  are  much  worse  than  " Vitamine." 


HISTORICAL  SURVEY 

In  spite  of  the  fact  that  the  knowledge  of  the  vitamines  taken  as 
a  whole  is  not  older  than  ten  years,  and  although  until  lately  the 
idea  was  prevalent  that  for  the  complete  nutrition  of  an  animal 
organism  only  proteins,  fats,  carbohydrates,  salts  and  water  were 
necessary,  there  is  nevertheless,  in  the  older  literature,  no  lack  of 
statements  which  of  themselves  should  have  given  rise  to  an  eager 
search  for  additional  dietary  components  essential  to  life.  The 
progress  which  has  already  been  made  by  research  in  vitamines 
removes  all  doubt  as  to  the  actual  existence  of  such  substances,  and 
every  year  brings  forth  new  findings  which  enhance  the  importance 
of  the  vitamines  to  life. 

The  scientific  research  leading  to  the  conception  of  the  vitamines 
proceeded  through  many  intermediate  stages,  which  we  shall  shortly 
describe.  A  great  stimulus  to  the  development  of  the  modern  science 
of  nutrition  was  furnished  by  the  investigations  into  the  chemistry 
of  the  proteins  which  we  owe,  above  all,  to  the  classical  work  of 
Emil  Fischer  and  Kossel.  These  investigations  not  only  contributed 
to  the  knowledge  of  the  composition  of  the  proteins,  but  also  gave  rise 
to  the  study  of  the  relationship  between  the  individual  dietary  con- 
stituents. They  demonstrated  particularly,  that  the  various  proteins 
exhibited,  qualitatively  as  well  as  quantitatively,  a  varied  composi- 
tion, and  that  naturally  occurring  proteins  have  not  the  same  physi- 
ological value.  Without  going  into  details,  which  may  be  found  in 
any  text-book,  we  shall  take  up  only  those  facts  bearing  upon  the 
study  of  the  vitamines.  It  has  been  shown  by  many  investigations 
that  certain  amino  acids,  such  as  tyrosine,  tryptophane,  arginine  and 
lysine,  are  more  or  less  indispensable  to  the  animal  organism.  This 
question  is  not  yet  completely  settled,  but  we  know,  however,  that 
some  proteins,  for  example,  zein  (which  is  lacking  in  tryptophane), 
or  gelatine  (which  lacks  several  important  amino  acids),  are  not 
sufficient  for  normal  maintenance  and  growth.  Latest  developments 
also  indicate  that  an  animal  will  utilize  a  diet  containing  animal 
proteins,  better  than  one  made  up  of  plant  proteins.  This  conception 
is  based  preeminently  upon  the  supposition  that  animal  proteins 
have  a  composition  more  nearly  related  to  body  protein  than  do  plant 

19 


20  THE    VIT AMINES 

proteins.  That  is  to  imply  that  the  animal  organism  needs  a  smaller 
amount  of  animal  protein  to  maintain  its  nitrogen  balance.  This 
view  was  utilized  by  Thomas  (1)  to  group  the  various  proteins  of 
animal  and  plant  origin  according  to  their  biological  value.  All 
these  questions  have  exerted  their  influence  upon  the  development 
of  the  study  of  the  vitamines,  and  are  even  now  closely  related  to 
many  of  our  problems  to  which  we  shall  call  attention  later. 

Another  matter  that  is  of  interest,  is  the  extent  of  the  ability  of 
the  animal  body  to  synthesize  many  of  the  substances  necessary  for 
its  existence.  The  peculiar  deficiency  symptoms,  of  which  we  speak 
in  this  book,  have  often  been  attributed  to  the  apparent  inability  of 
animals  to  synthesize  some  of  the  body  constituents.  The  sub- 
stances which,  of  all  others,  come  to  mind  here  are  the  nucleins  and 
the  lipoids  or  phosphatides.  If  this  were  really  the  case,  then  these 
substances  would  also  have  to  be  regarded  as  vitamines,  a  conception 
which  is  upheld  even  now  by  some  authors.  The  older  animal 
experiments,  conducted  with  purified  food  substances,  showed  that 
nucleins  and  lipoids  actually  could  be  synthesized.  In  spite  of  this, 
we  encounter  in  the  work  of  the  last  ten  years,  the  observation  that 
these  substances  have  exhibited  real  therapeutic  and  nutritive 
qualities,  an  observation  that  found  its  chief  exponent  in  H. 
Schaumann  (2)  who  elaborated  the  theory  of  phosphorus  insuffi- 
ciency as  an  explanation  of  beriberi  and  similar  diseases.  This  inter- 
pretation was  vigorously  opposed  by  the  writer  and  gave  him  the 
chance  to  propound  the  theory  of  the  vitamines.  We  shall  speak  of 
the  nucleins  and  lipoids  in  greater  detail  in  another  chapter,  but  we 
shall  also  touch  upon  them  lightly  at  this  point. 

Even  before  the  vitamines  were  known,  reports  were  published 
dealing  with  the  synthesis  of  lipoids  in  the  body.  McCollum, 
Halpin  and  Drescher  (3)  working  with  chickens  on  a  lipoid-free 
diet,  showed  that  the  eggs  contained  a  normal  amount  of  lecithin. 
Fingerling  (4)  and  also  Abderhalden  (5)  were  able  to  show  that 
animals  could  build  up  the  needed  organic  phosphorus  compounds 
from  inorganic  phosphates.  Still,  we  find  in  the  description  of  the 
condition  of  the  above  animals,  some  observations  that  point  to  the 
necessity  of  lipoids  for  life.  It  remained  for  vitamine  research  to 
clear  up  this  matter  completely. 

In  this  connection,  we  shall  mention,  first  of  all,  the  important 
work  of  Stepp  (6)  in  1909.  He  made  the  significant  observation  that 


HISTORICAL   SURVEY  21 

mice  cannot  maintain  themselves  on  a  diet  composed  of  bread,  and 
milk  which  has  been  extracted  with  alcohol.  If,  however,  the 
extracted  portion  is  recombined  with  the  diet,  then  the  animals 
begin  to  grow  once  more.  In  another  communication,  Stepp  (7) 
was  convinced  that  his  findings  were  not  due  to  the  loss  of  salts 
during  extraction,  and  classified  in  the  lipoid  group,  those  substances 
essential  to  life.  In  spite  of  the  fact  that  this  conclusion  was  not 
quite  justified,  the  work  of  Stepp  really  merited  more  attention  than 
it  received  at  the  time  of  its  publication.  Unfortunately,  this  was 
not  the  case,  for  the  relegation  of  the  substances  necessary  for  life 
to  the  class  of  lipoids,  which  already  had  been  considered  essential, 
did  not  signify  real  progress. 

Aside  from  the  work  of  Stepp,  attention  could  easily  have  been 
focused  upon  the  conception  of  the  vitamines  by  research  which  had 
for  its  purpose  the  study  of  the  importance  of  salts  in  nutrition. 
Relative  to  this,  the  school  of  Bunge  is  credited  with  a  great  deal  of 
merit.  Another  series  of  investigations  dealt  with  the  utilization  of 
purified  foodstuffs,  especially  in  the  mouse,  the  rat  and  the  dog. 

In  1873,  Forster  (8)  tried  to  determine  whether  or  not  dogs  could 
maintain  themselves  on  an  ash-free  diet.  For  this  purpose,  he  fed 
the  meat  remaining  after  the  preparation  of  Liebig's  meat  extract. 
These  residues  were  washed  repeatedly  with  distilled  water  until 
they  contained  only  0.8  per  cent  ash.  They  were  then  combined 
with  fat,  sugar  and  starch,  and  fed  to  dogs,  with  the  result  that  they 
died  sooner  than  starving  animals.  Experiments  with  pigeons,  fed 
with  casein  and  starch  (and  occasionally  a  little  fat),  gave  the  same 
results.  The  symptoms  noted  by  Forster,  in  pigeons,  were  partic- 
ularly interesting.  They  refused  food,  lost  a  great  deal  of  weight, 
showed  weakness,  opisthotonos  and  characteristic  circular  motions. 
As  we  shall  see  later,  these  symptoms,  which  Forster  attributed  to  a 
lack  of  salts,  were  apparently  identical  with  "  poly  neuritis  galli- 
narum,"  a  disease  of  chickens  described  by  Eijkman  in  1897.  This 
view  is  further  strengthened  when  we  note  the  length  of  time  that 
the  pigeons  lived — 13,  26  and  31  days.  In  dogs,  there  was  noted 
tremor,  peculiar  gait  and  weakness  similar  to  that  of  paresis. 

The  investigations  from  Bunge's  laboratory,  which  are  mentioned 
in  his  book  on  physiological  and  pathological  chemistry  (9),  are 
worthy  of  attention.  Lunin  (10)  reported  experiments  with  mice, 
fed  on  casein,  fat  and  cane  sugar.  Out  of  five  animals  used,  one 


22  THE    VITAMINES 

lived  11  days;  the  others,  13,  14,  15  and  21  days,  respectively,  while 
starving  animals  lived  only  from  3  to  4  days.  The  addition  of  soda, 
which  served  the  purpose  of  neutralizing  the  sulphuric  acid  arising 
from  the  protein  cleavage  prolonged  the  life  of  the  animals  to  a 
certain  extent;  the  ash  of  milk  likewise  had  the  same  effect.  Lunin 
explained  the  results  as  being  due  to  a  lack  of  organic  phosphorus 
compounds  (lecithin),  and  to  a  disturbance  in  the  balance  between 
the  inorganic  and  organic  food  components.  Subsequently,  Lunin 
made  the  unusually  important  observation  that  mice  could  thrive 
very  well  on  milk  powder  even  after  two  and  a  half  months,  and 
therefore  concluded  that  milk  contained  besides  the  known  elements, 
other  unknown  substances  essential  to  life.  That  this  conclusion 
had  already  been  arrived  at  in  1881  must  appear  to  us  as  truly 
remarkable.1  Socin  (11),  working  in  the  same  laboratory,  also  came 
to  a  similar  conclusion  in  1891.  His  problem  required  him  to  find 
out  whether  inorganic  or  organic  iron  is  the  better  utilized  in  the 
animal  body.  Mice  were  kept  on  a  diet  consisting  of  blood  serum, 
fat,  sugar,  starch,  cellulose  and  ash  (obtained  from  milk).  The 
missing  iron  was  added  in  the  form  of  hemoglobin,  hematin,  or  iron 
chloride,  without  the  least  effect  being  apparent  on  the  duration  of 
life.  All  of  the  animals  died  after  32  days,  while  the  controls,  fed 
on  yolk  of  egg,  starch  and  cellulose,  were  still  alive  after  99  days. 
Bunge  (12)  himself  obtained  the  same  results.  Hausermann  (13), 
working  with  rats,  guinea  pigs,-  rabbits,  cats  and  dogs,  kept  on  white 
bread  and  rice,  could  see  an  improvement  after  the  addition  of  iron, 
although  a  normal  state  of  nutrition  could  not  be  brought  about. 
The  experiments  of  Socin  were  confirmed  by  Hall  (14)  and 
Coppola  (15). 

Considering  simple  feeding  experiments,  we  find  the  work  of 
Pasqualis  (16)  on  chickens,  receiving  a  diet  composed  of  14  per  cent 
protein,  65  per  cent  corn  starch,  4  per  cent  dextrin,  4  per  cent  sugar, 
9  per  cent  olive  oil,  2  per  cent  wood  chips,  1  per  cent  common  salt 

1  Lunin  concluded  that  "mice  can  live  well  under  these  conditions  when 
receiving  suitable  foods  (milk)  but  as  the  above  experiments  demonstrate 
that  they  were  unable  to  live  on  proteins,  fats,  carbohydrates,  salts  and  water, 
it  follows  that  other  substances  indispensable  for  nutrition  must  be  present 
in  milk  besides  casein,  fat,  lactose  and  salts."  This  conclusion  was  quoted 
in  Bunge's  Physiological  and  Pathological  Chemistry,  but  did  not  attract 
the  attention  it  merited. 


HISTORICAL   SURVEY  23 

and  1  per  cent  salt  mixture  (in  the  form  of  corn  ash).  To  his  aston- 
ishment be  observed  that  the  animals  lost  weight  and  died — in  spite 
of  the  large  food  intake.  The  first  exact  feeding  experiments  on 
rats  were  carried  out  by  Henriques  and  Hansen  (17).  The  experi- 
ments were,  in  the  main,  of  short  duration,  lasting  only  from  three 
to  four  weeks.  During  this  time,  the  rats  gained  weight  and  showed 
a  retention  of  nitrogen,  but  the  experimental  period  was  apparently 
too  short  to  discover  any  food  deficiency.  Different  findings,  how- 
ever, were  reported  by  Falta  and  Noeggerath  (18)  who  repeated 
these  experiments.  They  fed  various  proteins  of  animal  origin 
together  with  fat,  sugar,  starch,  salts,  lecithin  and  cholesterol.  It 
was  possible  to  determine  the  value  of  the  various  proteins,  although 
not  one  of  the  combinations  was  permanently  sufficient,  as  was 
apparent  by  the  poor  nourishment  to  be  noted  after  varying  intervals 
of  time.  The  deficiencies  of  the  diet  made  their  appearance  only 
after  prolonged  feeding.  If  the  experiment  had  been  terminated  at 
the  end  of  four  weeks,  the  false  conclusion  could  have  been  made 
that  the  diet  had  been  adequate.  The  food  intake  was  diminished, 
and  the  interesting  observation  was  made  that  the  eyes  of  the  rats 
exhibited  pathological  changes,  of  which  we  shall  have  more  to  say 
later.  Working  in  the  laboratory  of  O.  Frank,  Jacob  (19)  reported 
some  feeding  experiments  with  pigeons  and  rats  on  a  diet  which  at 
that  time  was  thought  to  be  sufficient.  The  pigeons  died  after  a 
maximum  of  four  weeks  with  symptoms  of  severe  digestive  dis- 
turbance, although  they  lived  somewhat  longer  when  casein  was 
replaced  by  meat  powder.  Rats,  on  the  contrary,  lived  longer  on 
casein  and  died  after  43,  73  and  125  days.  Undoubtedly,  the  cause 
of  death  was  the  unsuitable  composition  of  the  diet,  though  Jacob 
considered  it  to  be  due  to  the  uniformity  and  lack  of  stimulating  sub- 
stances in  the  diet.  The  same  trend  of  thought  is  often  met  with 
in  the  older  as  well  as  the  newer  literature.  For  instance, 
McCollum  (20)  explained  the  dietary  deficiency  of  synthetic  diets 
as  being  due  to  lack  of  flavoring  and  stimulating  substances, 
although  at  present  he  is  the  most  eager  advocate  of  the  new 
ideas.  Through  the  addition  of  these  flavoring  and  stimulating 
substances,  he  was  unable  to  demonstrate  an  improvement  in  the 
nutritive  condition.  A  very  interesting  investigation  was  that  made 
by  Watson  (21).  He  fed  rats,  both  young  and  full  grown,  on  various 
meat  products.  The  young  animals  either  died  very  quickly  or 


24  THE    VITAMINES 

showed  inhibition  of  growth,  while  full  grown  animals  suffered  from 
paresis  and  sterility.  This  last  symptom  was  not  to  be  attributed 
to  lack  of  tryptophane.  The  animals  whose  growth  had  been  stunted 
resumed  normal  growth  on  a  mixed  diet. 

About  1906,  there  appeared  the  classical  work  of  Hopkins  and  his 
pupils,  who  displayed  marked  clearness  in  their  course  of  reasoning. 
Hopkins  fed  mice  on  a  mixture  which  contained  zein  (one  of  the  corn 
proteins)  which  lacks  tryptophane.  The  young  animals  were  able  to 
live  only  16  days,  whereas  upon  the  addition  of  this  amino  acid,  life 
was  prolonged  for  14  days  more.  Tyrosine  exerted  no  influence.  It 
appeared  from  these  results  that  the  above  diet  was  lacking  in  some- 
thing else  besides  tryptophane.  The  appearance  of  animals  so  fed, 
as  described  by  Wilcock  and  Hopkins  (22)  was  not  very  bright.  The 
animals  were  torpid,  had  cold  extremities,  half-closed  eyes  and  slimy 
fur.  At  that  time,  Hopkins  thought  that  tryptophane  could  be 
.conceived  of  as  the  precursor  of  adrenaline.  The  chemical  nature  of 
the  missing  components  (even  after  the  addition  of  tryptophane)  was 
not  recognized  at  that  time,  although  the  experiments  led  Hopkins  (23) 
to  the  prophetic  statement  which  is  reproduced  here  verbatim: 

,  But  further,  no  animal  can  live  upon  a  mixture  of  pure  protein,  fat  and 
carbohydrate,  and  even  when  the  necessary  inorganic  material  is  carefully 
supplied,  the  animal  still  cannot  flourish.  The  animal  body  is  adjusted  to- 
live  either  upon  plant  tissue  or  other  animals  and  these  contain  countless- 
substances  other  than  the  proteins,  carbohydrates  and  fats.  Physiological 
evolution,  I  believe,  has  made  some  of  these  well  nigh  as  essential  as  are  the 
basal  constituents  of  diet;  lecithin  for  instance,  has  been  repeatedly  shown 
to  have  a  marked  influence  upon  nutrition,  and  this  just  happens  to  be  some- 
thing familiar,  and  a  substance  that  happens  to  have  been  tried.  The  field 
is  almost  unexplored,  only  it  is  certain  that  there  are  many  minor  factors  in 
all  diets  of  which  the  body  takes  account.  In  diseases,  such  as  rickets,  and 
particularly  scurvy,  we  have  had  for  long  years  knowledge  of  the  dietetic 
factor,  but  though  we  know  how  to  benefit  these  conditions  empirically,  the 
real  errors  in  the  diet  are  to  this  day  quite  obscure.  They  are,  however, 
certainly  of  the  kind  which  comprises  these  minimal  quantitative  factors 
that  I  am  considering.  Scurvy  and  rickets  are  conditions  so  severe  that  they 
force  themselves  upon  our  attention,  but  many  other  nutritive  errors  affect 
the  health  of  individuals  to  a  degree  most  important  to  themselves,  and  some 
of  them  depend  upon  unsuspected  dietetic  factors. 

All  that  Hopkins  says  in  this  short  paragraph  applies  to  this  very 
day,  although  from  the  point  of  view  of  that  time  it  was  held,  for 
example,  that  lecithin  was  essential  for  life.  These  assertions  of 


HISTORICAL   SURVEY  25 

Hopkins  were  unknown,  because  he  offered  no  experimental  evidence 
in  their  support  till  1912.2 

Till  a  short  time  ago,  there  was  no  lack  of  opinions  which  denied 
the  existence  of  substances  essential  to  life.  We  have  only  to  recall 
the  work  of  Abderhalden  and  Lampe  in  1913  (25),  although  lately 
Abderhalden  has  changed  his  view  upon  the  matter.  He  (26)  con- 
ducted many  feeding  experiments  with  various  proteins,  hydrolyzed 
meat,  etc.,  without  once  recognizing  that  these  diets  were  lacking  in 
something  that  was  essential.3 

It  is  true  that  the  experiments  did  not  last  very  long,  and  that  the 
various  constituents  were  not  carefully  purified.  Incidentally,  it 
must  be  noted  that  in  these  experiments  short  periods,  in  which  a 
more  natural  diet  was  given,  were  frequently  interspersed  in  order 
to  abate  for  a  time  the  .distressing  deficiency  symptoms.  Above  all 
things,  we  must  recognize  that  in  dogs,  the  symptoms  manifested 
because  of  a  deficient  diet,  are  not  so  pronounced  as  in  pigeons  and 
rats.  Some  time  after  the  vitamine  theory  had  already  become 
familiar,  Abderhalden  (29)  found  indeed  that  digested  casein  had  less 
value  for  growing  animals  than  digested  meat.  He  immediately 
assumed  that  glycine  was  the  cause  of  the  difference,  whereupon  he 
started  various  experiments,  adding  this  amino  acid,  but  naturally 
without  any  success.  Similar  criticism  may  be  made  of  the  work  of 

2  For  instance,  Rohmann  (24)  is  mistaken  when  he  asserts  that  the  ideas 
expressed  by  us  originated  with  Hopkins.     Inasmuch  as  we  entered  the  field 
in  1911,  the  statements  of  Hopkins  were  unknown  to  us. 

3  While  I  was  working  in  the  city  hospital  at  Wiesbaden,  I  carried  out  under 
Abderhalden's  direction,   some  metabolism    experiments   with    dogs.     The 
animals  were  fed  with  edestin,  gliadin,  milk  powder  and  meat,  together  with 
other  usual  dietary  constituents.     Since  the  experiments  were  considered 
unsuccessful,  only  a  small  part  of  the  work  was  published  by  Abderhalden 
and  myself  (27).    At  that  time,  most  of  my  dogs  died,  so  that  the  feeding 
experiments  had  to  be  started  anew  a  number  of  times.    Even  then,  I  made 
the  observation  that  animals  fed  on  edestin  or  gliadin  quickly  recovered  on 
addition  of  milk  powder  or  meat.     I  wrote  to  Abderhalden  a  number  of  times, 
that  the  edestin  and  gliadin  combination  was  not  sufficient  to  make  the  animals 
thrive.    The  symptoms  which  I  observed  then  recall  those  noted  by  Chit- 
tenden  and  Underhill  (28)  in  dogs  fed  on  peas.     To  my  letters,  I  received  the 
na'ive  reply  from  Abderhalden,  that  in  such  metabolism  experiments,  success 
depends  upon  the  manner  in  which  the  animals  are  taken  care  of.     I  conscien- 
tiously attempted  to  follow  this  advice,  but  the  dogs,  on  such  a  poor  diet, 
refused  to  manifest  any  friendship  for  me. 


26  THE    VITAMINES 

Grafe  (30)  who  studied  the  protein-sparing  action  of  ammonium 
salts.  The  same  thing  applies  to  the  work  of  many  others  on  animal 
nutrition. 

One  of  the  last  opponents  of  the  vitamine  theory  was  the  late 
Rohmann  (31  and  I.e.  24).  He  expressed  the  idea  that  the  existence 
of  the  vitamines  was  seized  upon  only  because  certain  investigators 
had  had  poor  results  with  their  experimental  animals.  The  subject 
was  treated  by  Rohmann  in  a  rather  illogical  manner,  since  in  the 
end  he  admitted  the  existence  of  the  vitamines,  under  a  different 
name.  His  statements,  in  particular,  have  been  discussed  by  so 
many  authors  that  we  shall  not  consider  them  further  here. 

In  spite  of  the  great  powers  of  observation  possessed  by  such 
investigators  as  Bunge  and  Hopkins,  the  conception  of  the  vitamine 
theory  could  never  have  attained  its  present  importance,  if  a  powerful 
impulse  had  not  been  received  from  clinical  sources.  The  findings 
mentioned  above  have  not  been  utilized  in  human  pathology,  since 
most  clinicians  are  not  very  much  disposed  to  apply  the  results  of 
animal  experiments  to  man.  However,  great  interest  in  vitamines 
was  justifiably  aroused  when  the  applicability  of  the  results  to  human 
pathology  and  physiology  was  demonstrated.  For  a  long  time,  long 
before  the  publication  of  the  laboratory  findings,  the  literature  con- 
tained views  upon  the  etiology  of  scurvy,  rickets  and  pellagra  which 
appeared  to  be  very  nearly  correct.  The  conditions  surrounding 
these  diseases  were,  however,  too  complicated  to  be  suitable  for  a 
direct  experimental  research.  On  the  other  hand,  as  regards  beriberi, 
the  circumstances  were  entirely  different.  Here  we  had  to  deal  with 
a  problem  the  etiology  of  which  was  comparatively  simple,  for  the 
disease  could  be  brought  logically  into  causal  relationship  with  the 
continued  consumption  of  rice.  Nevertheless,  many  years  of  effort 
were  necessary  for  these  conceptions  to  gain  a  foothold  in  the  litera- 
ture. When  the  first  edition  of  this  book  was  being  written  in  1913, 
it  was  still  necessary  for  us  to  wage  a  hard  fight  in  support  of  our 
contention.  Such  is  not  the  case  at  present,  for  the  characterization 
of  beriberi  as  an  avitaminosis  has  met  with  general  recognition.  At 
the  end  of  this  chapter,  we  shall  discuss  the  history  of  beriberi 
research,  since  it  may  logically  serve  as  an  introduction  to  the  study 
of  the  vitamines. 

We  shall  begin  here  with  a  discussion  of  scurvy,  although  this 
disease  really  did  not  contribute  any  direct  stimulus  to  vitamine 


HISTORICAL   SURVEY  27 

research.  Scurvy,  however,  is  the  first  disease  the  etiology  of  which 
was  associated  with  a  definite  mode  of  nutrition.  The  reasons  why 
scurvy  gave  no  immediate  impulse  to  research  were  to  be  looked  for 
in  the  variety  of  the  feeding,  which  might  have  been  responsible  for 
the  onset  of  the  disease.  It  was  difficult  to  conceive  of  the  disease 
as  being  due  to  a  lack  of  one  and  the  same  substance  iii  the  diet. 

It  was  Kramer  (32),  an  Austrian  army  physician,  who  recognized 
for  the  first  time,  the  existence  of  scurvy.  In  1720  with  a  field  army 
in  Hungary,  he  was  confronted  with  a  severe  epidemic  of  scurvy.  He 
wrote  to  the  authorities  and  to  his  colleagues  to  secure  help. 
A  shipment  of  dried  antiscorbutic  herbs  was  hurried  to  him  in  spite 
of  which  thousands  died  of  this  disease.  He  then  made  the  following 
entry  in  his  book: 

Scurvy  is  a  terrible  disease  for  which  there  is  no  known  cure.  Medication 
does  not  help,  neither  does  surgery.  Be  careful  of  bleeding;  shun  mercury 
as  a  poison.  The  gums  may  be  massaged,  the  stiff  joints  may  be  rubbed 
with  fat — but  all  in  vain.  If  one  could  only  have  available  a  supply  of  green 
vegetables,  or  a  sufficient  amount  of  the  vital  antiscorbutic  juices;  or  if  one 
could  have  at  hand  oranges,  limes  or  lemons,  or  their  preserved  pulp  or  juice 
so  that  a  lemonade  could  be  made  out  of  them;  or  administered  as  such  in 
three  or  four  ounce  doses — then  one  could  be  in  a  position  to  cure  this  dreadful 
disease,  without  other  help. 

As  we  see,  Kramer  selected  a  method  of  treatment  which  could 
not  be  better  chosen  even  today. 

Bachstrom  (33)  recognized  in  1734  that  the  incidence  of  scurvy 
was  not  due  to  cold  weather,  sea  air  or  salted  meat,  but  to  a  lack  of 
fresh  vegetables.  The  latter,  he  perceived,  was  the  primary  cause 
of  the  disease.  Lind  (34),  in  his  work  on  scurvy,  has  noted  many 
cases  which  were  cured  by  administration  of  oranges  or  lemons. 
Cider  was  next  to  oranges  in  its  efficacy.  At  that  time,  he 
made  the  important  observation  that  very  severe  cases  could  be 
cured  in  6  days,  and  recognized  also  that  hard  work  accentuated  the 
symptoms  of  scurvy.  Coming  now  to  the  modern  history  of  scurvy, 
we  must  give  prominence,  above  all  others,  to  the  name  of  Barlow  (35) 
who  attributed  the  onset  of  infantile  scurvy  (also  called,  M  oiler- 
Barlow's  disease)  to  milk  which  had  been  heated  for  a  long  time.  It 
was  only  lately,  in  1907,  that  scurvy  in  guinea  pigs  was  discovered — 
a  discovery  of  the  greatest  significance  to  vitamine  research.  The 
work  of  Hoist  and  Frolich  (36)  on  this  subject  was*  repeated  on  all 
sides,  and  their  results  were  completely  corroborated. 


28 


THE   VITAMINES 


Research  on  rickets  was  undertaken  experimentally  only  in  late 
years,  although  Miller  (37)  had  already  associated  rickets  with  a 
particular  dietary  even  before  the  vitamines  were  known.  He 
thought  that  many  of  his  cases  were  due  to  a  lack  of  butter,  and 


FIG.  1 


FIG.  2 


FIG.  3 


FIG.  1.    DECREASE  IN  THE  NUMBER  OF  CASES  OF  BERIBERI  IN  THE  JAPANESE 
NAVY  AFTER  THE  INTRODUCTION  OF  MEAT  INTO  THE  DIETARY  (TAKAKI) 

FIG.    2.  THE  CALORIC  VALUE  OF  THE  FOOD  DIMINISHED  (TAKAKI)  (CF.  FIG.  1) 
FIG.  3.  THE  BODY  WEIGHT  INCREASED  (TAKAKI)  (CF.  FIGS.  1-2) 

consequently  prescribed  milk,  cream,  butter,  egg-yolk  and  cod  liver 
oil.  Schabad  (38)  saw  a  marked  difference  between  olive  oil,  sesame 
oil,  and  cod  liver  oil  in  their  influence  on  rickets,  and  in  a  later 
work  (39)  he  discusses  the  question  as  to  the  nature  of  the  curative 
substance. 


HISTOEICAL   SURVEY 


29 


He  was  convinced  that  it  was  not  a  ferment  and  that  its  efficacy 
was  not  diminished  after  heating  for  one  hour  at  100°C.  Since  the 
war,  great  strides  have  been  made  in  our  knowledge  of  scurvy  and 
rickets,  and  we  shall  discuss  them  in  later  chapters. 

Beriberi  is  an  avitaminosis  about  which  we  are  best  informed. 
Although  known  for  hundreds  and  perhaps  thousands  of  years,  it  is 
only  in  the  last  25  years  that  actual  progress  has  been  made  relative 
to  its  occurrence  and  prevention.  This  is  perhaps  due  to  the  increase 
in  the  number  of  cases  of  beriberi,  coincident  with  the  introduction 
of  modern  machinery  for  rice  milling.  It  was  quite  properly  pre- 
sumed by  Wernich  (40)  and  van  Leent  (41)  that  there  existed  a 
causative  relationship  between  rice  consumption  and  beriberi.  In 
1882,  Takaki  (42)  proposed  a  change  in  the  rice  diet  of  the  Japanese 
navy,  so  that  meat,  bread,  fruit  and  vegetables  were  added.  Since 
that  time  beriberi  has  almost  completely  disappeared. 

Takaki's  tables  are  exceptionally  instructive.  It  is  apparent  from 
Fig.  1,  that  upon  the  elimination  of  the  rice  diet  in  1882,  there  was 
an  immediate  drop  in  the  number  of  cases  of  beriberi,  which  number 
subsequently  remained  trifling.  In  Figs.  2  and  3  it  may  be  seen 
further,  that  although  the  caloric  value  of  the  new  diet  was  smaller 
in  comparison  to  that  of  the  previous  one  (causing  beriberi),  there 
was  a  definite  increase  in  the  average  weight  of  the  men.4 

Even  greater  strides  were  made  in  Java  by  Dutch  investigators. 
Based  upon  very  rich  statistical  material  collected  at  Eijkman's 
suggestion  by  Vordermann  (43)  in  a  large  number  of  Javanese  prisons, 
it  was  possible  to  demonstrate  that  the  disease  was  associated  with 
the  ingestion  of  white  (polished)  rice.  The  fluctuation  in  the  number 
of  cases  of  beriberi  due  to  various  kinds  of  rice  is  well  illustrated  in 
Vordermann's  Table: 


NUMBER   OP 

NUMBMR   OF 

BERIBERI  CASES 

PRISONERS 

CASES 

KATI°  NUMBER  OP  PRISONERS 

White  rice  

150,266 

4,201 

1:39 

Rice  with  partial  "silver  skin".  . 
Unpolished  rice 

35,082 
96  530 

85 
9 

1:416 
1:10725 

The  findings  of  Vordermann  were  corroborated  by  Braddon  (44) 
on  a  large  number  of  cases  in  the  Malay  peninsula.    He  was  able  to 


4  Figures  1,  2,  and  3. 


30  THE    VITAMINES 

show  that  some  natives,  like  the  races  of  Tamils,  who  live  on  " cured" 
or  "parboiled"  rice,  remain  free  from  this  disease.  This  rice  was  so 
prepared  that  it  was  steamed  before  using;  the  husk  came  off  easily, 
thus  obviating  the  necessity  for  polishing.  To  convince  himself  of 
the  accuracy  of  Braddon's  observations,  Fletcher  (45)  in  1905  under- 
took, in  the  Kuala  Lumpur  insane  asylum,  some  research  on  lunatics, 
which  lasted  for  one  year.  His  purpose  was  to  determine  the  differ- 
ence between  " cured"  and  ordinary  decorticated  rice.  Throughout 
the  entire  perod,  the  diet  varied  only  qualitatively;  out  of  120 
patients  on  polished  rice,  36  contracted  beriberi  from  which  18  died, 
while  from  123  patients  on  " parboiled"  rice,  only  two  developed 
beriberi,  and  these  two  cases  were  admitted  as  such  into  the  hospital. 
In  1909,  Ellis  (46),  working  at  the  insane  asylum  in  Singapore, 
reported  similar  investigations,  which  extended  back  as  far  as  1901. 
In  his  fruitful  statistics,  he  showed  definitely  that  the  number  of 
beriberi  cases  decreased  steadily  from  year  to  year,  upon  gradually 
substituting  steamed  rice  for  polished  rice.  Analagous  results  were 
also  obtained  on  healthy  laborers  by  Fraser  (47) . 

In  1897,  Eijkman  (48)  (cf.  Vordermann),  found  that  beriberi  is 
brought  about  by  a  long  continued  consumption  of  white  rice,  since 
polishing  removes  a  substance  which  is  protective  against  the  out- 
break of  beriberi.  The  Dutch  investigators  were  of  the  opinion 
that  the  most  important  part  of  the  rice  grain,  the  so-called  "silver- 
vlissen"  (silver  skin),  could  neutralize  the  toxins  of  white  rice.  There- 
upon, it  fell  to  Fraser  and  Stanton  (49)  to  make  clear  the  point  that 
upon  polishing  rice,  more  than  just  the  "silver  skin"  is  lost. 

Eijkman's  (50)  discovery  of  experimental  beriberi  marked  a  great 
step  forward.  This  finding  was  made  accidentally  since  Eijkman 
observed  that  chickens  which  fed  upon  the  remains  of  the  food  used 
in  a  hospital  for  beriberi  died  of  a  disease  which  he  recognized  oppor- 
tunely to  be  similar  to  human  beriberi  This  discovery,  which  was 
made  in  1896  (the  disease  was  called  "polyneuritis  gallinarum"), 
made  it  possible  to  get  away  from  experiments  upon  man,  and  assured 
the  collection  of  more  valuable  experimental  data  in  a  shorter  time. 

After  the  discovery  of  experimental  beriberi,  Eijkman  went  a 
step  further.  He  found  that  the  addition  of  the  pericarp  of  the  rice 
kernel,  or  even  the  rice  bran,  to  white  rice  made  it  possible  to  prevent 
the  occurrence  of  beriberi  in  animals.  For  these  observations, 
however,  he  did  not  find  the  correct  explanation ;  he  believed  that  the 


HISTORICAL   SURVEY  31 

starch  of  the  grain  gave  rise  to  toxins  which  exerted  a  deleterious 
action  on  the  nervous  system,  and  that  this  was  prevented  by  the 
addition  of  the  pericarp.  This  conception  should  not  surprise  us 
since  in  those  days  the  nature  of  the  disease  was  quite  puzzling. 
Eijkman  (51)  likewise  made  the  important  observation  that  the 
watery  extract  of  rice  bran  possessed  therapeutic  properties.  Phytin 
was  found  in  the  rice  bran,  but  it  was  shown  to  be  without  effect  on 
beriberi.  He  noted  also  that  the  curative  substance  is  dialysable 
and  not  precipitated  by  the  addition  of  alcohol.  The  observations 
made  in  1897  were,  after  all,  those  upon  which  the  modern  research 
on  beriberi  is  based.  All  credit  is  due  Eijkman  for  having  laid  the 
foundation  for  the  conduct  of  future  experiments. 

Grijns  (52),  continuing  the  work,  was  able  to  confirm  the  experi- 
ments of  Eijkman  completely.  He  was  the  first  worker  to  express 
clearly  a  conception  of  beriberi,  which  holds  good  to  this  day.  Grijns 
said  that  the  disease  developed  when  the  diet  was  lacking  certain 
substances  which  were  of  importance  in  the  metabolism  of  the 
peripheral  nervous  system.5  Curative  substances  similar  to  those 
occurring  in  rice  bran  were  found  by  Grijns  in  a  kind  of  bean  called 
"Katjang-idjoe"  (Phaseolus  radiatus),  and  in  meat;  he  also  demon- 
strated that  these  foodstuffs  lost  their  curative  properties  when  they 
were  heated  to  120°C.  These  experiments  were  of  greatest  signifi- 
cance in  the  further  development  of  the  question,  and  they  were 
also  confirmed  by  Eijkman  (I.e.  51).  Breaudat  (54)  used  rice  bran 
in  the  treatment  of  human  beriberi  with  good  results.  Eraser  and 
Stanton  (55)  sought  to  determine  more  definitely  the  nature  of  the 
substance.  They  found  that  it  was  soluble  in  strong  alcohol  and 
that  it  retained  its  activity  after  the  removal  of  alcohol-soluble 
proteins.  They  made  analyses  of  various  kinds  of  rice  and  believed 
that  a  rice  poor  in  phosphorus  would  cause  beriberi.  Thereupon, 
they  suggested  the  phosphorus  content  as  a  -practical  indicator 
of  the  nutritive  value  of  rice.  For  instance,  rice  which  contained 
a  minimum  of  0.46  per  cent  P2O5  was  to  be  considered  harmless. 
However,  we  should  not  forget  that  Schiiffner  and  Kuenen  (56) 
have  shown  that  the  method  of  preparation  of  the  rice  diet  is 
likewise  of  importance.  That  is,  the  rice  should  be  partaken  of 
together  with  the  broth,  particularly  in  the  case  of  whole  rice. 

5  Eijkman  carried  out  a  lengthy  polemic  on  this  point,  but  he  admits  now 
(53)  that  Grijns  was  correct. 


32  THE    VITAMINES 

If  the  broth  is  regularly  discarded  then  beriberi  can  easily  occur, 
even  with  whole  rice.  Based  upon  the  findings  of  Eraser  and  Stanton, 
Schaumann  (57)  thought  of  beriberi  as  a  disturbance  in  metab- 
olism due  to  a  lack  of  organic  phosphorus  compounds.  This  theory 
was  likewise  applied  to  other  avitaminoses,  such  as  scurvy  and  ship 
beriberi.  It  found  disciples  (Simpson  and  Edie  (58)),  and  dominated 
this  field  of  pathology  till  the  advent  of  the  vitamine  theory.  At 
that  time,  it  was  quite  plausible  to  regard  the  great  difference  in 
phosphorus  content  between  white  rice  and  rice  bran,  and  the 
undeserved  acclaim  of  organic  phosphorus  compounds,  as  thera- 
peutic factors  in  medicine. 

In  the  years  that  followed,  there  appeared  in  quick  succession  a 
great  number  of  experiments  dealing  with  the  chemical  nature  of  the 
curative  substance.  Various  foodstuffs  were  used  for  this  purpose. 
Hulshoff  Pol  (59)  showed  that  a  watery  extract  of  "Katjang-idjoe" 
beans,  clarified  with  lead  acetate,  contained  the  curative  substance. 
From  this  clear  filtrate,  Pol  obtained  a  crystalline  substance  which 
he  called  "X-acid;"  there  is  a  lack  of  further  information  as  to 
whether  this  substance  possessed  any  characteristic  physiological 
properties. 

Schaumann  (I.e.  2),  who  extended  the  list  of  curative  substances 
to  include  yeast  (which  was  used  by  Thompson  and  Simpson  (60)  in 
the  treatment  of  human  beriberi),  investigated  the  influence  of  the 
already  known  yeast  constituents.  Among  these  he  investigated 
the  influence  of  yeast  nucleic  acid  and  yeast  lecithin,  but  without 
obtaining  definite  results.  Eijkman  (61)  showed  that  the  active 
substance  of  yeast  could  be  extracted  with  88  per  cent  alcohol.  Funk 
(62)  had  already  noticed  this  before,  but  by  this  method  only  an 
incomplete  extraction  could  be  effected.  Teruuchi  (63)  extracted  rice 
bran  with  dilute  hydrochloric  acid,  neutralized  the  solution,  thereby 
precipitating  phytiri,  then  evaporated  the  filtrate  and  extracted  the 
residue  with  alcohol.  This  extract  was  active  and  contained  only  a 
small  proportion  of  its  original  phosphorus  content.  Similar  results 
were  also  obtained  by  Chamberlain  and  Vedder  (64).  They  found 
that  the  curative  substance  was  adsorbed  with  animal  charcoal  and 
tried  to  develop  a  procedure  based  upon  this  finding,  but  failed. 
Shiga  and  Kusama  (65)  found  that  the  active  principle  of  rice  bran 
was  destroyed  by  heating  to  130°C.  with  0.5  per  cent  hydrochloric 
acid  or  with  1  per  cent  soda  solution,  but  not  at  100°C. 


HISTORICAL   SURVEY  33 

Research  on  the  preparation  of  the  active  substance  from  rice 
polishings  were  also  conducted  by  Tsuzuki  (66),  but  with  little  suc- 
cess. Owing  to  the  enormous  content  of  phytin  in  rice  bran,  Aron 
and  Hocson  (67)  believed  that  it  was  curative;  the  good  results  they 
obtained  may  be  explained  by  the  probability  that  the  phytin  was 
contaminated  by  some  of  the  active  substance.  Research  on  phytin 
had  already  been  carried  out  by  Eijkman  without  results,  and  also 
by  Cooper  and  Funk  (68). 

To  summarize  our  knowledge  of  the  chemical  nature  of  the  active 
principle  prior  to  the  introduction  of  the  vitamine  theory  (till  1911), 
the  following  may  be  set  down  with  certainty: 

1.  The  substance  is  soluble  in  water,  alcohol  and  acidified  alcohol. 

2.  The  substance  is  dialysable. 

3.  The  substance  is  destroyed  at  130°C. 

When  we  took  up  the  question  in  1911,  it  was  not  known  whether 
the  active  substance  was  organic  or  inorganic  in  nature,  whether  or 
not  it  was  a  constituent  of  proteins,  nucleins  or  phosphatides.  It  was 
not  certain  that  we  were  not  dealing  with  a  ferment,  nor  was  it  known 
if  the  substance  belonged  to  some  chemical  group  already  described, 
or  to  some  new  unknown  class  of  substances.  We  shall  be  in  a 
position  to  answer  a  good  many  of  these  questions  during  the  course 
of  our  discussions. 


INTRODUCTORY 

In  the  historical  part,  we  have  seen  that  the  animal  organism 
cannot  live  very  long  upon  an  artificially  prepared  diet.  Since 
systematic  investigations  into  the  vitamine  requirements  of  all  classes 
of  plants  and  animals  have  not  yet  been  made,  it  is  not  possible  to 
say  with  entire  certainty  that  the  above  contention  is  generally  true. 
The  firmly  established  importance  of  the  vitamines  for  the  existence 
of  certain  animals  and  plants,  organisms  which  are  far  removed  from 
each  other  genetically,  makes  it  apparent  that  these  substances  are 
of  universal  importance  to  life. 

It  is  evident  that  there  are  differences  in  the  qualitative  and 
quantitative  requirements  and  it  may  later  develop  that  various 
organisms  need  various  quantities  as  well  as  various  kinds  of  vita- 
mines.  We  shall  briefly  enumerate  the  facts  that  have  led  us  to  this 
conclusion,  and  point  out  how  many  types  of  vitamines  are  known 
up  to  the  present.  We  choose  to  speak  of  vitamine  types  in  this 
case,  and  not  of  definite  vitamines,  for  as  long  as  these  substances 
remain  unidentified,  and  till  such  time  as  they  may  be  compared 
with  each  other  in  the  pure  state,  it  is  obviously  impossible  to  talk 
of  their  identity.  Until  now,  we  have  differentiated  three  such 
types.  Whether  with  these  three  types  we  have  finally  reached 
the  possible  limit,  it  cannot  be  stated  definitely;  however,  it  appears 
unlikely  that  Nature,  in  the  variety  of  its  manifestations,  essential 
conditions  and  intensity  of  metabolism,  should  limit  itself  to  onjy 
three  types.  On  the  other  hand,  it  is  possible  that  after  purification, 
that  which  seemed  to  us  a  single  substance  might  well  prove  to  be 
a  mixture.  We  may  be  dealing  with  complex  substances  having 
some  chemical  groups  in  common,  and  others,  the  significance  of 
which  has  not  yet  been  demonstrated. 

It  would  be  quite  premature  to  propose,  with  McCollum  (69),  that 
only  two  or  three  vitamines  exist  in  nature.  He  arrived  at  this 
conclusion  as  a  result  of  his  numerous  rat  experiments,  which  showed 
that  for  this  species  two  types  of  vitamines  were  sufficient,  and 
thereupon  drew  conclusions  that  were  to  be  applied  to  the  entire 
plant  and  animal  kingdom.  Although  we  are  very  well  informed  as 
to  the  food  requirements  of  rats,  compared  to  those  of  other  organ- 

35 


36  THE    VITAMINES 

isms,  we  cannot  maintain  that  we  know  all  that  there  is  to  be  known 
of  the  requisite  food  constituents,  as  long  as  we  shall  not  have 
obtained  them  (including  vitamines)  in  the  chemically  pure  form. 
This  applies  naturally  even  more  to  other  animals,  about  whose 
metabolism  we  are  still  less  informed.  It  is  therefore  of  greatest 
importance,  at  this  stage  of  our  knowledge,  to  disregard  generaliza- 
tions of  this  sort  since  they  are  detrimental  to  the  development  of 
the  questions  that  interest  us. 

We  shall  now  consider  the  work  leading  to  the  present  conception 
of  the  three  different  vitamine  types.  When  we  pointed  out  in  1912 
(I.e.  62) l  that  the  animal  organism  needs  vitamine  for  complete 
nutrition,  the  term,  "vitamine,"  was  used  only  in  a  very  general  way. 
At  that  time,  we  drew  a  distinction  between  the  antiberiberi,  anti- 
scorbutic and  antirachitic  vitamines,  although  this  classification  was 
based  only  upon  the  physical  characteristics,  origin  and  influence  on 
metabolism.  New  facts  have  been  brought  to  light,  but  this  classi- 
fication has  remained  unchanged.  We  knew  in  1912  that  the  pericarp 
of  grains  contained  something  that  was  of  importance  in  the  metab- 
olism of  man  and  certain  species  of  birds.  Then  Schaumann  (I.e.  2) 
found  that  yeast  and  some  animal  extracts  contained  a  subtance  of 
similar  nature.  Furthermore,  it  was  known  that  man  as  well  as 
the  guinea  pig  was  in  need  of  a  vitamine  as  a  protection  against 
scurvy — a  vitamine  that  was  recognized  by  us  to  be  quite  different 
from  the  antiberiberi  vitamine.  In  spite  of  the  above  mentioned 
facts  and  the  work  of  Stepp  (I.e.  6,  7)  already  referred  to,  and  in 
spite  of  the  results  of  vitamine  research  in  1911,  the  new  ideas  were 
not  immediately  applied  to  the  general  science  of  nutrition.  The 

1  I  regarded  it  of  paramount  importance,  that  the  then  ruling  conception 
of  the  necessity  of  the  lipoids  or  the  nuclein  substances  was  substituted  by  the 
fundamentally  different  vitamine  theory.  At  the  same  time,  I  must  admit 
that  when  I  chose  the  name,  " vitamine,"  I  was  well  aware  that  these  sub- 
stances might  later  prove  not  to  be  of  an  amine  nature.  However,  it  was 
necessary  for  me  to  choose  a  name  that  would  sound  well  and  serve  as  a  catch- 
word, since  I  had  already  at  that  time  no  doubt  about  the  importance  and 
the  future  popularity  of  the  new  field.  As  we  have  noted  in  the  historical 
part,  there  was  no  lack  of  those  who  suspected  the  importance  of  still  other 
dietary  constituents,  besides  those  already  known,  for  the  nutrition  of  animals. 
These  views  were  unfortunately  unknown  to  me  in  1912,  since  no  experimental 
evidence  had  appeared  in  their  support.  I  was,  however,  the  first  one  to 
recognize  that  we  had  to  deal  with  a  new  class  of  chemical  substances,  a  view 
which  I  do  not  need  to  alter  now  after  eight  years. 


INTKODUCTORY  37 

only  investigation  on  this  subject  was  the  classical  work  of  Hopkins 
(70)  which  appeared  in  1912,  demonstrating  that  the  addition  of  a 
small  quantity  of  milk  to  an  artificial  diet  induced  growth  in  rats. 
The  amount  of  milk  was  so  small  as  to  be  negligible,  as  far  as  its 
energy  factor  was  concerned.  In  these  experiments,  there  was  no 
attempt  to  differentiate  between  the  various  vitamines,  since  milk 
contains  all  that  is  necessary  to  life.  While  the  vitamine  conception 
had  at  that  time  attained  a  definite  standing  in  England  (not  without 
vigorous  effort),  the  dissemination  of  these  ideas  in  other  countries 
met  with  but  poor  success.  For  instance,  Abderhalden  and  Lampe 
denied  the  existence  of  vitamines  in  1913,  and  Rohmann  even  in 
1916.  In  the  United  States  there  appeared  simultaneously  with  the 
beginning  of  vitamine  research  the  very  important  work  of  Osborne  and 
Mendel  (71)  on  the  artificial  feeding  of  rats,  which  greatly  advanced 
our  knowledge  of  the  food  value  of  the  various  kinds  of  proteins. 
These  investigations  were  indeed  the  first  to  be  carried  out  with 
such  carefully  purified  proteins  and  over  such  a  long  period  of  time 
(more  than  one  year,  one-third  of  the  lifetime  of  a  rat).  These 
experiments  served  two  purposes,  first,  to  determine  the  nutritional 
value  of  various  proteins,  and  secondly,  to  determine  how  long  rats 
can  live  on  an  artificial  diet.  At  that  tune,  we  emphasized  the  fact 
that  for  such  investigations  it  is  particularly  important  to  provide 
the  animals  with  vitamines,  if  clear  results  are  to  be  obtained. 
Besides  this,  we  showed  that  in  many  nutrition  experiments  the  diet 
was  in  some  unknown  manner  contaminated  by  vitamines  which 
were  responsible  for  the  length  of  time  the  animals  survived.  This 
was  later  shown  to  be  true;  for  example,  we  could  demonstrate  the 
presence  of  some  nitrogen  -  containing  impurity  in  milk  sugar;  the 
same  is  true  of  other  products  derived  from  milk.  In  the  experi- 
ments of  Osborne  and  Mendel,  the  diets,  taken  as  a  whole,  were 
obviously  lacking  in  vitamines,  since  many  of  the  animals  died 
suddenly,  or  they  would  have  died  if  the  diet  had  not  been  changed 
quickly.  Young  animals  lived  for  a  certain  time,  but  mostly  failed 
to  grow. 

In  another  investigation,  Osborne  and  Mendel  (72)  described 
experiments  in  which  rats  were  fed  on  a  mixture  that  could  have 
been  thought  of  as  fat-free.  It  contained,  among  other  things, 
protein-free  powdered  milk  extracted  with  ether.  In  this  case  also, 
normal  growth  was  obtained.  The  same  authors  then  analyzed  the 


38  THE    VITAMINES 

protein-free  milk  for  its  inorganic  constituents,  and  prepared  an 
artificial  protein-free  milk  having  the  same  composition;  this  also 
permitted  of  good  growth.  In  a  later  communication,  these  inves- 
tigators (73)  pointed  out  that  the  results  were  not  so  favorable  with 
the  artificial  preparation.  Their  animals  lived  from  114  to  277  days, 
but  after  this  they  died  without  the  post-mortem  giving  any  plausible 
reason  for  death.  In  one  case  particularly,  the  addition  of  natural 
food  was  ineffective  in  preventing  death. 

Hopkins  and  Neville  (74)  reported  that  they  attempted  to  replace 
milk  by  a  preparation  made  according  to  Osborne  and  Mendel. 
They  stated  that  when  they  used  purified  lactose,  which  had  been 
prepared  from  milk,  the  animals  failed  to  grow.  McCollum  and 
Davis  (75)  found  that  rats,  weighing  40  to  50  grams,  could  grow 
normally  for  about  three  months  on  the  Osborne-Mendel  diet.  At 
the  end  of  this  time  the  animals  stopped  growing,  but  were  in  good 
general  health.  In  the  light  of  our  work  on  vitamines,  the  authors 
came  to  the  conclusion  that  the  failure  to  grow  was  not  due  to  lack 
of  salt,  fat  and  phosphatides,  but  to  lack  of  vitamines.  They  found 
the  necessary  substances  in  egg-yolk  and  butter,  especially  in  the 
ether-soluble  portion.  Besides  this,  they  found  that  rats  on  the 
above  diet  produced  very  little  milk,  and  that  the  young  were 
stunted.  Normal  growth  was  restored  upon  the  addition  of  the 
ether  extract  of  eggs  or  butter. 

Almost  simultaneously  with  the  work  of  McCollum  and  Davis, 
there  appeared  the  paper  of  Osborne  and  Mendel  (76)  in  which  they 
confirmed  their  earlier  findings.  As  a  result  of  these  studies,  they 
showed  that  artificial  milk,  like  the  natural  protein-free  variety,  is 
not  sufficient  for  growth.  To  obtain  adequate  growth,  either  milk 
powder  or  butter  had  to  be  added. 

In  later  publications,  McCollum,  as  well  as  Osborne  and  Mendel, 
disregarded  the  importance  of  the  antiberiberi  vitamine  for  growth 
more  and  more,  and  reiterated  the  existence  of  a  specific  growth 
substance  in  certain  fats,  like  butter,  egg-yolk  and  others.  Little 
by  little,  the  entire  vitamine  structure  became  shaky,  especially  after 
Osborne  and  Mendel  (77)  had  started  their  studies  on  butter.  By 
centrifugation,  they  separated  butter  into  three  fractions.  They 
reported  that  the  pure  butter  fat,  supposedly  nitrogen  and  phosphorus 
free,  still  retained  its  activity.  In  particular,  they  were  able  to  cure 
ophthalmia  (keratomalacia,  a  disease  of  which  we  shall  speak 


INTRODUCTORY  39 

later)  in  rats  kept  on  an  artificial  diet.  These  experiments  were 
intended  to  show  that  the  growth  substance  could  not  be  a  vitamine. 
We  do  not  yet  know  the  chemical  nature  of  the  substance  in  butter, 
but  there  is  nothing  to  disprove  the  conception  that  this  substance 
contains  nitrogen  and  may  be  classified  as  a  vitamine,  especially 
when  it  is  remembered  in  what  small  amounts  it  may  be  active.2 

Working  with  Macallum  (81)  on  the  fractionation  of  butter,  we 
demonstrated  that  when  we  followed  the  procedure  of  Osborne  and 
Mendel,  and  dissolved  large  amounts  of  butter  fat  (about  12  kilos) 
in  acetone  and  shook  out  this  solution  with  dilute  hydrochloric  acid, 
we  found  23.4  mgm.  of  nitrogen  in  the  extract;  after  hydrolysis  of 
the  fatty  residue  with  dilute  hydrochloric  acid,  an  additional  22  mgm. 
was  obtained.  It  is  quite  obvious  that  it  is  impossible  to  remove 
all  the  nitrogen  from  butter  with  the  above  procedure.  In  spite  of 
these  findings,  it  is  true  that  butter  is  poor  in  nitrogen  and  therefore 
it  must  contain  a  substance  that  is  active  in  minimal  amounts. 
McCollum  and  Davis  (82)  endorsed  this  opinion,  inasmuch  as  they 
doubted  the  absence  of  nitrogen  in  butter.  In  order  to  answer  the 
question,  Osborne  and  Wakeman  (83)  conducted  another  experiment 
with  butter  and  found  that  it  contained  traces  of  nitrogen  and 

*  2  Later  on,  McCollum  and  Kennedy  (78)  sought  to  introduce  the  classi- 
fication, "water-soluble  B"  for  the  antiberiberi  vitamine  and  "fat-soluble 
A"  for  the  antirachitic  vitamine;  lately,  Drummond  (79)  used  the  term  "water- 
soluble  C"  for  the  antiscorbutic  vitamine.  Regarding  this  matter,  I  have 
attempted  to  show  (80)  that  these  designations  are  incorrect,  chemically 
and  logically.  It  would  be  totally  inaccurate  to  differentiate  substances 
extracted  from  complicated  mixtures  by  their  solubility  in  certain  solvents. 
It  would  seem  to  be  of  importance  to  replace,  in  the  English  publications, 
such  designations  as  "growth-promoting,  water-soluble,  accessory  B  factor," 
by  something  more  simple.  For  this  purpose,  I  suggest,  at  least  for  the  present, 
the  adoption  of  the  following  nomenclature : 

Vitamine  B  for  the  antiberiberi  vitamine 
Vitamine  A  for  the  antirachitic  vitamine 
Vitamine  C  for  the  antiscorbutic  vitamine 

These  letters  are  already  used  very  often  in  the  English  literature,  and  the 
nomenclature  to  be  used  in  this  book  is  suggested  as  being  simple  and  tune 
saving,  particularly  in  indexing  the  literature.  Since  this  has  been  written, 
a  note  by  Drummond  (80a)  has  appeared  in  which  the  same  kind  of  nomen- 
clature is  proposed,  except  that  he  suggests  "vitamin"  instead  of  "vitamine" 
for  the  English  scientific  literature.  I  cannot  agree  to  this  change  since 
I  still  believe  in  the  nitrogenous  nature  of  these  substances. 


40  THE   VITAMINES 

phosphorus.  The  question  as  to  the  nitrogen  content  of  butter  has 
riot  been  touched  upon  for  some  years,  since  the  solution  of  the 
problem  could  only  be  obtained  through  a  purification  of  vitamine  A. 
In  this  connection,  the  amount  of  lecithin  in  butter,  based  upon  the 
phosphorus  content,  was  calculated  to  be  0.017  per  cent  by  Wrampel- 
meyer  (84)  and  from  0.04  to  0.07  per  cent  by  Supplee  (85) ;  the  latter 
also  demonstrated  the  presence  of  choline,  trimethylamine,  and 
ammonia. 

While  Osborne  and  Mendel,  as  well  as  McCollum,  regarded  butter 
and  certain  other  fats  as  the  only  growth-promoting  substances, 
Funk  and  Macallum  (86)  went  to  the  opposite  extreme,  stating  that 
vitamine  B  was  the  only  growth-promoting  substance.  It  appears 
from  the  newer  investigations,  as  we  have  always  believed,  that 
vitamine  B  is  far  more  important  to  life  and  is  required  in  greater 
amounts  then  vitamine  A.  Nevertheless,  both  substances  are 
necessary  for  growth  as  has  been  developed  by  the  above  controversy. 
The  Funk-Macallum  experiments  showed  that  butter,  when  added 
to  an  artificial  diet,  does  not  cause  growth,  and  it  appeared  that  an 
addition  of  yeast  was  imperative.  To  be  sure,  there  was  then  no 
apparent  difference  in  the  efficacy  of  butter  and  lard,  but  it  should 
be  said  here  that  in  our  investigations  we  found  it  necessary  to  add 
a  large  amount  of  yeast  in  order  to  obtain  good  results;  fresh  yeast 
worked  better  than  dried  yeast.  It  is  not  impossible  that  yeast,  in 
the  fresh  condition,  contains  some  vitamine  A;  on  the  other  hand,  in 
the  light  of  the  results  obtained  by  Daniels  and  Loughlin  (87),  it  is 
possible  that  some  fats,  till  now  looked  upon  as  vitamine-free,  contain 
enough  of  this  vitamine  to  stimulate  growth  in  rats. 

In  this  manner,  the  existence  of  two  vitamines,  A  and  B,  was 
definitely  established.  In  addition,  it  became  apparent  that  the 
substance  playing  the  greatest  part  in  the  growth  of  rats  is  either 
identical  with  the  antiberiberi  vitamine  or  belongs  to  the  same  type. 
Coming  now  to  vitamine  C,  its  individuality  and  its  differentiation 
from  the  other  two  vitamines  was  determined  in  the  following 
manner.  While  we  find  vitamine  C  often  associated  with  vitamine  B 
in  Nature,  there  are  products  such  as  egg-yolk  and  cereals,  which 
are  very  poor  in  vitamine  C  but  especially  rich  in  vitamine  B.  The 
evidence  of  their  dissimilarity  may  be  seen  more  clearly  in  the  work 
of  Seidell  (88) ,  who  showed  that  vitamine  B  could  be  adsorbed  quan- 
titatively with  Lloyd's  reagent  (fuller's  earth).  Harden  and  Zilva 


INTRODUCTORY  41 

(89)  then  showed  that  from  a  mixture  of  autolyzed  yeast  and  orange 
juice,  substances  frequently  used  as  a  source  of  both  vitamines, 
vitamine  B  could  be  completely  removed  by  adsorption  with  fuller's 
earth,  while  the  presence  of  vitamine  C  could  be  demonstrated  in  the 
filtrate.  These  findings  have  been  corroborated  by  Byfield,  Daniels 
and  Loughlin  (90)  incidental  to  some  other  work.  Vitamine  C  is  the 
most  unstable  of  the  vitamines,  although  it  appears  to  be  more 
stable  in  juices  that  are  slightly  acid.  -In  association  with  these 
facts,  there  is  the  discovery  of  Fiirst  (91),  who  was  the  first  one  to 
show  that  if  grains  are  allowed  to  germinate  a  formation  of  vitamine 
C  takes  place,  which  disappears  again  upon  drying  and  reappears 
upon  the  addition  of  water.  We  have  thought  (1.  c.  62)  that  in 
this  observation  there  was  a  possibility  of  finding  a  genetic  relation- 
ship between  vitamine  C  and  vitamine  B.  Through  progress  in  the 
methods  of  demonstrating  the  presence  of  vitamine  B,  it  may  be 
possible  to  investigate  the  question,  if  with  the  formation  of  vitamine, 
C  there  is  a  corresponding  diminution  in  vitamine  B.  The  relation* 
ship  between  these  vitamines  has  become  more  interesting,  since 
Osborne  and  Mendel  (92)  showed  that  known  antiscorbutics,  such 
as  fruits,  always  contain  a  definite  amount  of  vitamine  B.  Although 
we  know  that  this  vitamine  has  no  effect  upon  scurvy,  the  reverse  of 
this,  the  influence  of  vitamine  C  on  beriberi,  is  not  sufficiently  estab- 
lished. It  might  be  possible  for  instance,  that  for  the  cure  of  scurvy 
very  small  amounts  of  vitamine  C  are  necessary,  whereas  to  in- 
fluence beriberi  similarly  much  larger  doses  are  required.  Thiq 
matter  must  be  cleared  up  by  further  research. 

We  must  also  put  forth  the  question  whether,  with  the  enumeration 
of  the  above  three  vitamines,  we  have  exhausted  all  the  possibilities. 
There  is  no  definitive  answer  to  this  at  present,  but  it  is  frequently 
touched  upon  in  the  latest  investigations,  and  it  came  into  prominence 
through  a  review  of  the  available  data  by  Mitchell  (93),  pointing  to 
the  idea  that  vitamine  B  is  different  from  the  growth-promoting 
substance.  In  spite  of  this,  we  must  note  that  the  conclusions  of 
Mitchell  are  not  indisputable.  He  compared  earlier  work  done  with 
comparatively  inefficient  methods,  with  newer  work  that  is  more 
liable  to  be  accurate,  and  consequently  it  is  not  to  be  wondered  at 
that  he  found  a  difference;  besides,  he  compared  extracts  made  from 
different  materials.  Somewhat  later,  there  appeared  the  first  experi- 
ment on  this  matter  of  Emmett  and  Luros  (94),  which  at  least  had, 


42  THE   VITAMINES 

the  advantage  that  the  source  of  material  (unpolished  rice)  was  the 
same  in  the  comparison  of  both  characteristics.  We  shall  comment 
upon  the  questions  referred  to,  particularly  the  vitamines,  in  greater 
detail,  but  at  this  point  we  wish  to  emphasize  that  the  functions  of 
growth  and  the  curative  action  on  beriberi  were  tested  on  two  dif- 
ferent animals,  the  pigeon  and  the  rat.  Heated  unpolished  rice  still 
retained  its  growth  promoting  property,  although  to  a  somewhat 
lesser  degree,  while  its  influence  on  pigeon  beriberi  was  completely 
destroyed.  We  can  find  many  explanations  for  this  behavior  without 
necessarily  accepting  the  existence  of  two  vitamines  of  the  same  type. 
Above  all,  we  do  not  know  just  what  are  the  vitamine  requirements 
of  these  types  of  animals.  We  shall  see  furthermore  that  all  the 
known  facts  point  to  the  idea  that,  although  it  shall  have  been  demon- 
strated that  both  substances  differ  from  each  other,  they  would 
still  be  related  chemically.  In  this  connection,  it  might  be  shown 
that  various  types  of  animals  can  utilize  the  different  stages  of  vita- 
mine  cleavage  with  varying  degrees  of  success.  This  could  proceed,  on 
the  one  hand,  through  a  difference  in  synthetic  abilities  and,  on  the 
other,  through  certain  specific  symbiotic  intestinal  flora. 

A  similar  question  is  brought  up  in  reference  to  vitamine  A  (anti- 
rachitic  vitamine).  Mellanby  (95)  holds  the  view,  that  in  the 
etiology  of  rickets,  that  vitamine  which  according  to  our  nomen- 
clature must  be  called  antirachitic  vitamine  (vitamine  A)  is  of 
importance.  It  is  found  in  fats  such  as  butter,  egg-yolk,  and  cod 
liver  oil.  Whereas  Mellanby  worked  with  young  dogs,  Hess  (96), 
working  with  rachitic  children,  found  that  milk  fat  had  no  effect, 
while  cod  liver  oil  was  found  to  be  beneficial.  From  this,  Hess  drew 
the  conclusion  that  the  growth-promoting  vitamine  found  in  butter 
was  different  from  the  antirachitic  vitamine.  Here  also,  both  sub- 
stances belong  to  the  same  type,  and  we  must  bear  in  mind  the  main 
question  whether  the  vitamines,  like  other  natural  substances,  do 
not  also  deviate  from  each  other  somewhat  chemically,  in  spite  of  the 
possibility  that  they  play  the  same  or  very  nearly  the  same  role  in 
physiology. 

Much  more  important  and  interesting  are  the  experiments  which 
deal  with  the  part  played  by  milk  and  milk  products.  We  are 
sometimes  almost  tempted  to  assume  that  milk,  aside  from  the  vita- 
mines  already  mentioned,  contains  still  other  essential  substances. 
This  applies  not  only  to  milk,  but  also  to  products  made  from  milk, 


INTRODUCTORY  43 

such  as  casein,  lactalbumin,  and  partly  also  to  lactose.  Obviously 
another  conclusion  is  possible  here,  namely,  that  milk  contains  the 
most  suitable  combination  of  amino  acids  for  the  animal  body.  It 
was  pointed  out  by  McCollum  and  Davis  (97)  that  heating  of  casein 
for  one  hour,  at  one  atmosphere  in  an  autoclave,  materially  decreases 
its  food  value;  Funk  and  Macallum  (98),  attempting  to  confirm  this 
observation,  found  that  the  facts  were  somewhat  in  accord,  but  that 
the  diminished  nutritive  value  of  casein  could  be  remedied  by  the 
administration  of  fresh  orange  juice.3 

We  believed,  at  that  time,  that  heating  of  the  casein  destroyed  the 
adsorbed  C-vitamine,  and  our  view  was  strengthened  by  the  later 
work  of  Harden  and  Zilva  (100)  and  Drummond  (I.e.  79).  We  (101) 
had  already  been  able  to  show  that  beriberi  in  pigeons  could  be  cured 
by  the  addition  of  the  vitamine  fraction  of  lime  juice,  although  we 
were  not  certain  if  this  effect  was  not  due  to  the  presence  of  vita- 
mine  C.  Still  later,  Osborne  and  Mendel  (I.e.  92)  showed  that  the 
favorable  influence  of  orange  juice  on  the  growth  of  rats  was  due  to 
the  presence  of  vitamine  B.  This  observation  was  further  confirmed 
by  the  work  of  Byfield,  Daniels  and  Loughlin  (I.e.  90),  in  which  they 
showed  that  orange  juice  treated  with  fuller's  earth  exerted  no 
growth-promoting  influence  either  on  young  or  grown  rats,  in  spite 
of  still  containing  vitamine  C.  Till  now,  there  has  been  no  explana- 
tion of  why,  when  the  diet  already  contains  apparently  sufficient 
vitamine  B,  upon  the  addition  of  more  of  this  vitamine  the  organism 
should  respond  with  an  acceleration  of  growth.  There  is  a  possi- 
bility here  that  orange  juice  may  contain  a  new  vitamine  which, 
together  with  B  vitamine,  is  adsorbed  by  fuller's  earth. 

Osborne,  Wakeman  and  Ferry  (102)  showed  that  certain  types 
of  proteins,  for  instance,  edestin,  manifest  a  greater  capacity  for 
adsorbing  vitamine  B  than  do  other  proteins,  and  that  the  latter,  by 
means  of  thorough  washings,  could  not  be  freed  from  it.  It  would  be 
plausible  to  assume  that  casein,  as  well  as  lactalbumin,  might  show  a 
selective  adsorptive  capacity  for  some  new  vitamine.  As  we  shall 
show  towards  the  end  of  this  book,  the  acceptance  of  a  new  and 
important  amino  acid  or  of  a  new  vitamine  appears  inviting  for  the 
elucidation  of  the  etiology  of  certain  pathological  conditions,  and 
also  for  the  explanation  of  the  difference  in  food  value  between 
proteins  of  vegetable  and  animal  origin. 

3  Hogan  (99)  found  that  heating  of  proteins,  casein  in  particular,  does  not 
impair  their  effectiveness. 


44  THE    VITAMINES 

In  a  great  number  of  experiments,  Osborne  and  Mendel  (103) 
showed  that  the  nutritive  value  of  lactalbumin  is  much  greater  than 
that  of  casein,  and  they  attributed  it  to  the  nature  of  the  particularly 
favorable  amino  acid  composition  of  this  protein.4  Edelstein  and 
Langstein  (104)  recently  arrived  at  the  conclusion  that  the  superiority 
of  mother's  milk  over  cow's  milk  for  the  child  is  due  to  the  greater 
content  of  lactalbumin  in  mother's  milk. 

In  a  series  of  papers,  which  are  not  altogether  clear  in  their  mean- 
ing, Emmett  and  Luros  (105)  sought  to  show  that  while  lactalbumin 
of  itself  was  a  complete  protein  for  growth,  the  specially  favorable 
results  obtained  must  be  attributed  to  the  influence  of  the  protein- 
free  milk  added  at  the  same  time.  In  this,  they  coincide  with 
the  view  of  McCollum,  Simmonds  and  Parsons  (106).  In  other 
words,  the  favorable  influence  of  lactalbumin  on  the  growth  of  rats 
was  to  be  attributed  to  the  presence  of  a  vitamine  in  the  protein-free 
milk  or  the  lactose.  This  vitamine  was  supposed  to  be  different 
from  vit amines  B  and  A. 

Emmett  and  McKim  (107)  have  also  assumed  the  existence  of 
two  vitamines  in  yeast — one  that  cures  beriberi  and  one  that  pro- 
motes growth  in  animals.  They  reached  these  conclusions  from 
adsorption  experiments  with  fuller's  earth.  They  showed  that  the 
activated  fuller's  earth  was  protective  against  beriberi,  but  did  not 
influence  the  weight  of  the  animal.  Such  experiments,  unless  well 
controlled,  can  have  no  real  significance,  considering  the  possibility 
of  incomplete  adsorption,  the  relative  amounts  used  and  other 
factors. 

Sugiura  and  Benedict  (108)  point  out  that  certain  diets  with  the 
addition  of  yeast  serve  to  increase  the  growth  of  young  rats,  although 
the  mothers  appear  to  be  lacking  in  milk.  They  conclude  therefore 
that  for  this  purpose  a  special  vitamine  is  desirable.  As  an  illustra- 
tion of  this,  the  example  is  given  that  an  addition  of  casein  exerts  a 
much  greater  influence  than  a  simple  addition  of  extra  protein.  In 
one  particular  case  it  was  shown  that  purified  casein  could  not  be 
replaced  by  purified  meat.  In  a  second  experiment,  they  diminished 
the  amount  of  the  added  yeast  to  0.5  per  cent  of  the  total  diet  and  it 
therefore  appeared  not  impossible  that  the  failure  of  milk  production 

4  Later,  these  investigators  showed  that  this  is  true  only  when  protein-free 
milk  is  used  as  source  of  vitamine  B.  This  addition  seemed  to  have  the  effect 
of  completing  the  value  of  that  particular  protein. 


INTRODUCTORY  45 

was  due  to  an  insufficient  amount  of  vitamine  B.  Unfortunately, 
these  investigators  failed  to  determine  whether  an  addition  of  B- 
vitamine  would  not  stimulate  milk  secretion.  Invariably,  all  of  the 
above  mentioned  workers  stressed  the  possibility  of  finding  a  still 
unknown  vitamine,  especially  in  milk. 

In  conclusion  we  wish  to  mention  that  besides  other  possibilities 
of  clearing  up  the  etiology  of  pellagra,  we  have  already  intimated 
(I.e.  62)  that  there  might  be  a  specific  antipellagra  vitamine — a  view 
held  as  well  by  Goldberger  (109).  This  vitamine  could  be  a  sub- 
stance easily  adsorbed  from  animal  protein  as  we  pointed  out  on 
page  43.  Still,  for  these  or  for  other  conceptions  of  pellagra,  no 
real  experimental  evidence  is  at  present  available. 

After  having  discussed  the  definitely  established  types  of  vitamines, 
we  shall  take  up  the  vitamine  requirements  of  various  organisms. 


PARTI 

The  Vitamine  Requirements  of  Plants 
and  Animals 


CHAPTER   I 
THE  R6LE  OF  THE  VITAMINES  IN  THE  VEGETABLE  KINGDOM 

It  was  quite  clear  from  the  start  that  the  importance  of  the 
vitamines  was  not  limited  to  animal  life  only,  but  was  applicable 
to  all  living  matter.  At  present,  the  view  is  held  that  the  animal 
organism  does  not  possess  the  ability  of  synthesizing  these  factors, 
and  in  this  respect  it  is  dependent  either  directly  or  indirectly  on 
plant  life.  We  must  determine,  however,  which  plants,  and  espe- 
cially what  parts  of  plants,  possess  the  power  of  building  up  the 
vitamines  out  of  simpler  combinations.  Furthermore,  we  should 
like  to  know  what  role  these  substances  play  in  plant  physiology.  We 
can  immediately  assert  that  although  the  work  on  this  aspect  of  the 
subject  is  still  in  its  infancy,  not  all  plants  apparently  are  able  to 
build  up  the  vitamines  (this  is  perhaps  not  true  of  all  vitamines). 

Upon  more  careful  consideration  of  the  higher  plants,  we  find  the 
vitamines  localized  in  two  divisions,  in  the  seeds  and  in  the  green 
leaves,  parts  of  the  plant  which  at  certain  times  are  of  particular 
importance  in  its  metabolism.  We  find,  with  few  exceptions,  the 
greatest  amount  of  vitamine  B  in  seeds,  together  with  varying 
amounts  of  vitamine  A.  It  is  quite  apparent  that  in  seeds,  the 
analogue  of  milk  and  eggs  in  the  animal  kingdom,  all  the  substances 
are  present  which  are  necessary  for  the  sprouting  of  the  new  plant. 
The  vitamines  play  a  great  part  in  this  process,  yet  we  are  dealing 
here  only  with  conjectures.  As  soon  as  seeds  are  planted  under 
conditions  of  proper  moisture  and  temperature,  the  ferments,  which 
had  been  in  a  quiescent  state,  commence  to  function.  These  fer- 
ments begin  to  marshal  the  stored  up  reserve  substances.  In  this 
instance,  we  may  also  consider  the  vitamines  as  reserve  substances, 
and  we  may  conceive  that  they  too  undergo  some  chemical  changes 
through  the  influence  of  ferments.  As  an  analogy  to  this  example 
we  already  know  that  vitamine  C  is  generated  upon  the  germination 
of  seeds.  All  other  phenomena,  like  the  division  of  the  nuclei  of 
cells,  remain  unknown  to  us,  although  we  are  soon  aware  of  the 
appearance  of  the  first  green  shoot.  What  part  the  vitamines  play 
in  this  case  is  not  definitely  known.  Perhaps  they  are  of  importance 

49 


50  THE   VITAMINES 

in  the  first  stages  of  metabolism;  perhaps  they  are  responsible  for 
the  sudden  onset  of  cell  division.  In  organisms  which  multiply  by 
conjugation  or  copulation,  the  vitamines  or  similar  substances  exert 
some  influence  perhaps  in  the  subsequent  nucleus  and  protoplasm 
partition.  When  we  compare  the  quiescent  with  the  sprouting  seed, 
we  see  the  difference  between  the  living  and  the  dead  tissue,  although 
the  gross  chemical  compositions  in  both  conditions  are  not  far 
different  from  each  other.  The  entire  difference  may  be  due  to  the 
formation  of  an  active  vitamine  modification.  It  is  evident  that  no 
great  progress  is  made  when  an  unknown  matter  is  explained  by 
something  equally  unknown.  It  may  nevertheless  be  of  value,  as 
far  as  future  investigations  are  concerned,  to  point  out  in  which 
class  of  chemical  bodies  the  substances  essential  to  life  are  to  be 
sought. 

Clark  (110)  described  the  influence  and  characteristics  of  a  sub- 
stance which  he  called  "Oocytin,"  and  which  was  supposed  to  be 
able  to  bring  about  the  formation  of  the  fertilization  membrane  of 
sea  urchin  eggs,  previously  treated  with  strontium  chloride.  This 
substance  was  prepared  in  powdered  form  from  centrifuged  and 
defibrinated  ox  blood.  The  powder  gave  the  reaction  of  protein, 
purine,  and  pentose,  and  contained  only  a  small  amount  of  phos- 
phorus. It  was  inactivated  by  heating  to  73°  to  80°C.,  or  by  long 
contact  with  alcohol.  Similar  substances  were  demonstrated  in  the 
sperm  of  the  sea  urchin  by  Robertson  (111).  These  or  other  active 
substances  naturally  play  a  similar  part  in  the  plant  kingdom, 
although  they  are  for  the  moment  unavailable  for  physiological- 
chemical  work. 

The  question  as  to  the  ability  of  certain  types  of  plants  and  par- 
ticular parts  of  plants  to  build  up  vitamines  is  still  far  from  solution. 
In  higher  plants,  for  example,  there  are  three  possibilities: 

1.  In  seeds,    stored  up   vitamines   suffice   to   stimulate   further 
synthesis  of  vitamines  in  the  growing  parts  of  the  plants. 

2.  It  is  possible  that  the  small  amount  of  vitamines  in  seeds  suffices 
to  carry  the  plants  along  to  a  certain  stage,  from  which  there  is  a 
symbiosis  with  certain  microorganisms  which  provide  the  plants  with 
vitamines. 

3.  It  is  possible  that  some  of  the  plant  structures,  for  example, 
the  leaves,  possess  the  ability  to  synthesize  certain  vitamines. 


THE   VITAMINES   IN   THE   VEGETABLE    KINGDOM  51 

The  subject  of  this  chapter  serves  merely  as  a  suggestion  of  the 
important  role  of  the  vitamines  in  plant  life,  without  which  knowledge 
of  the  vitamines  can  never  be  complete.  The  very  few  facts 
mentioned  here  should  serve  to  stimulate  further  work  in  this 
direction. 

YEAST 

Since  the  discovery  of  its  curative  influence  on  beriberi  by  Schau- 
mann  (I.e.  2),  yeast  appears  to  be  the  best  material  available  for  the 
chemical  study  of  the  vitamines.  Of  all  natural  products,  it  seems 
to  be  the  one  most  rich  in  vitamine  B  and  therefore  is  worthy  of 
special  attention.  One  could  be  tempted  to  assume  that  since  yeast 
cells  are  so  rich  in  vitamines  and  possess  such  an  energetic  metab- 
olism of  their  own,  they  should  be  able  to  prepare  their  own 
vitamines.  This  is,  however,  only  partially  the  case,  as  has  been 
already  shown  in  some  older  neglected  work  which  has  recently 
been  repeated,  and  confirmed.  Pasteur  (112)  made  the  obser- 
vation in  1871  that  the  capacity  of  yeast-cell  growth  depended 
very  much  upon  the  size  of  the  inoculation.  He  found  also  that 
addition  of  certain  organic  substances  accelerates  the  fermenta- 
tion of  certain  types  of  yeasts.  It  was  Wildiers  (113)  who  correctly 
understood  Pasteur's  observation  and  developed  it  further.  It  may 
well  be  said  that  Wildiers  foresaw  the  existence  of  the  vitamines 
as  far  back  as  1901.  His  important  studies  have  demonstrated 
that  upon  the  addition  of  a  small  quantity  of  sterile  yeast  extract 
to  a  nutritive  solution,  containing  an  ammonium  salt  as  its  sole 
source  of  nitrogen,  a  small  inoculation  is  sufficient;  the  growth 
of  the  cells  being  measured  in  this  case  by  the  CO*  evolved.  He 
recognized  that  this  growth  stimulus  could  not  be  attributed  to 
the  presence  of  the  hitherto  known  factors,  and  named  this  new 
substance  "bios."  Its  characteristics  were  not  so  far  different 
from  that  which  we  have  come  to  know  as  B-vitamine.  It  was 
soluble  in  water  and  also  in  80  per  cent  alcohol,  and  insoluble  in 
ether.  It  was  dialyzable  and  could  not  be  precipitated  by  the  well- 
known  precipitants,  phosphotungstic  acid  included.  The  substance 
was  found  to  be  stable  in  acid  medium  but  on  heating  for  a  short 
time  with  alkali  it  was  destroyed.  The  investigation  of  the  resistant 
qualities  of  this  substance  gave  no  clear  results.  The  presence  of 
bios  was  never  shown  in  the  decomposition  products  of  egg  albumin 


52  THE   VITAMINES 

split  by  acids,  and  likewise  not  in  yeast  ash.  The  experiments  by 
Wildiers  were  then  confirmed  by  Amand  (114)  who  demonstrated 
that  the  failure  of  the  yeast  to  grow,  with  just  a  slight  inoculation, 
was  not  due  to  the  toxicity  of  the  nutritive  solution.  Devloo  (115) 
reported  that  he  isolated  pure  bios  from  lecithin.  From  his  own 
data  it  is  evident  that  his  preparation  was  not  very  active,  and  at 
present  we  can  say  with  certainty  that  the  activity  noted  was  due 
to  the  contamination  of  the  lecithin  with  vitamine.  In  this  connec- 
tion, it  has  been  also  stated  that  lecithin  is  curative  for  beriberi;  we 
shall  have  occasion  to  refer  to  this  matter  later. 

We  should  not  think  that  the  important  conclusions  of  Wildiers 
found  immediate  acceptance  in  the  literature.  For  example,  H.  H. 
Pringsheim  (116)  took  the  stand  that  the  bios  of  Wildiers  was  nothing 
else  than  protein  material,  which  was  best  used  in  the  form  that  is 
found  in  yeast  cells  themselves.  This  statement  of  Pringsheim's 
was  introduced  into  the  literature,  and  therefore  the  conclusions  of 
Wildiers  were  given  no  further  attention.  For  instance,  Rubner 
(117)  said  that  the  growth  of  yeast  began  only  when  there  was  a 
certain  relationship  between  the  amount  of  food  and  the  number  of 
yeast  cells,  an  excess  of  food  acting  as  a  stimulus  for  yeast  growth. 
From  time  to  time,  however,  reports  appear  in  the  literature  dealing 
with  the  observation  of  Wildiers.  Vlahuta  (118)  prepared  a  peptone 
from  beer  yejast,  with  cold  sufturic  acid,  which  could  give  rise  to 
fermentation.  Kurono  (119)  f  otmd  that  a  vitamine  extract,  prepared 
from  rice  polishings,  added  to  Hayduck  or  Nageli  solution,  accelerated 
to  a  greater  extent  the  growth  of  yeast  cells  and  also  the  degree  of 
fermentation,  than  when  peptone  or  asparagine  were  added.  Brill 
and  Thurlow  (120),  on  the  contrary,  could  obtain  no  increase  in  the 
growth  of  yeast  cells  with  rice  polishings.  The  ability  of  certain 
brewery  residues  to  accelerate  fermentation  can  be  attributed  to  the 
presence  of  vrtamines.  Moufang  (121)  made  the  observation  that 
dead  beer  or  baker's  yeast  exerted  a  marked  catalytic  action  on 
fermentation.  Saito  (122)  was  of  the  opinion  that  certain  chemical 
substances  were  important  to  allow  of  a  complete  development  of  the 
reproductive  capacity  of  yeast.  Bokorny  (123)  observed  an  increase 
in  weight  of  yeast  when  it  was  grown  on  urine  containing  sugar 
(we  sliall  see  later  that  urine  contains  vitamine).  Vansteenberge 
(124)  studied  the  influence  of  autolyzed  yeast  upon  the  growth  of 
yeast  and  lactic  acid  bacteria,  and  found  that  above  a  certain 


THE    VITAMINES    IN   THE    VEGETABLE    KINGDOM 


53 


optimum  concentration,  this  autolysate  is  no  longer  active  and  even 
inhibits  somewhat  the  growth  of  the  cells — a  finding  that  is  of  greatest 
importance  for  our  arguments.  He  found  later  that  the  autolyzate 
must  be  diluted  to  obtain  the  best  results.  Ceucine,  tyrosine,  and 
asparagine  have  an  inhibiting  influence  on  the  growth  of  yeast  cells 
but  not  on  the  lactic  acid  bacteria.  Recently  Lampitt  (125)  has 
shown  that  the  increase  in  the  quantity  of  yeast  cells  depends  on  the 
original  number  present.  Evidently,  this  observation  is  to  be 
explained  in  this  way :  that  due  to  excessive  inoculation,  a  number  of 
dead  cells  are  implanted  at  the  same  time  and  that  the  living  cells 
can  utilize  the  vitamine  set  free. 

Since  the  growth  of  yeast  cells  might  be  utilized  as  a  means  for 
the  determination  of  antiberiberi  vitamine,  systematic  investigations 
were  undertaken  during  the  last  two  years  for  the  purpose  of  studying 
the  vitamine  requirements  of  these  cells.  It  was  expected  in  this 
way  to  measure  vitamine  B,  but  we  shall  presently  show  that  this 
is  not  the  case.  In  spite  of  the  possibility  that  the  substance  pro- 
moting the  growth  of  yeast  belongs  to  the  type  of  B-vitamine, 
newer  work  has  shown  that  the  substance  promoting  the  growth 
of  yeast  is  not  identical  with  vitamine  B.  It  is  possible,  however, 
that  for  yeast,  an  organism  characterized  by  greater  synthetic 
ability,  the  cleavage  products  of  vitamines  might  be  sufficient, 
whereas  in  higher  animals,  on  the  contrary,  the  entire  vitamine 
complex  must  be  administered. 

In  1912,  the  author,  using  the  fermentation  method,  endeavored 
to  show  that  vitamine  could  act  as  a  co-ferment,  but  without  success. 
Abderhalden  and  Schaumann  (126)  were  apparently  more  fortunate 
and  they  described  a  phosphorus-free  substance,  obtained  from 
yeast,  which  could  favorably  influence  the  fermentation  of  certain 
sugars.  Abderhalden  and  Kohler  (127)  likewise  described  the 
growth-promoting  influence  of  some  yeast  fractions  on  the  yeast 
cells.  The  same  authors  also  investigated  the  influence  of  yeast 
extracts  on  growth  of  Flagellata  (Colpoda)  and  Algae  (Ulothrix) 
with  positive  results.  A  whole  series  of  experiments  on  this  sub- 
ject has  lately  been  published  in  the  United  States.  The  method 
of  R.  J.  Williams  (128)  denoted  progress  in  that  it  substituted  for 
fermentation  a  direct  observation  on  the  growth  of  yeast  cells. 
The  method  is  as  follows:  A  hanging  drop  is  prepared  so  that  it 
contains  the  least  number  of  yeast  cells  (one  if  possible)  obtained  by 


54  THE   VITAMINES 

inoculating  Nageli's  solution  with  a  needle-point  of  yeast  culture, 
preparing  at  the  same  time  the  necessary  controls.  The  cells  are 
counted  after  5  to  6  hours  and  again  after  20  to  24  hours.  From 
20  to  several  thousands  of  cells  could  develop  from  a  single  cell  in 
24  hours,  depending  upon  the  vitamine  concentration.  Williams 
undertook  some  problems  with  the  help  of  this  method,  for  example, 
the  protective  influence  of  fat  upon  the  destruction  of  vitamines  by 
alkali.  Vitamine  extracts,  prepared  with  acid  or  alkali,  appeared 
to  be  best  utilized  by  yeast.  The  greatest  influence  was  obtained  by 
a  preparation  made  by  shaking  yeast  with  fuller's  earth.  The  ac- 
tive substance  was  also  shown  to  be  present  in  the  alcoholic  extract 
of  protein-free  milk,  wheat  embryo,  and  pancreatin,  and  in  the 
decomposed  phosphotungstic  acid  fraction  from  yeast.  Casein 
and  lactose  were  likewise  active,  although  in  the  former  the  ac- 
tivity was  not  attributed  to  the  amino  acids  but  to  the  small  amounts 
of  vitamines  present.  The  active  substance  was  partly  destroyed 
by  heating  in  the  autoclave  for  a  half  hour  at  one  atmosphere.  An 
alcoholic  extract  of  egg-yolk  was  active  while  the  ether  extract 
containing  vitamine  A  was  inactive.  The  C-vitamine,  as  we  shall 
note  later,  appears  to  have  no  influence  on  the  growth  of  yeast. 
Bachmann  (129)  investigated  the  behavior  of  two  kinds  of  yeast, 
one  apparently  "  Saccharomyces  cerevisiae"  and  the  other,  isolated 
from  fermented  pears.  These  two  varieties  showed  a  different 
behavior  towards  the  vitamine  solutions.  One  of  the  yeasts  grew  on 
the  surface  and  was  less  dependent  upon  the  vitamine  addition  (in 
this  she  indicated  the  possible  analogy  of  green  leaves  and  influence 
of  oxygen).  The  Bachmann  method  is  as  follows:  A  yeast  sus- 
pension in  a  liquid  nutritive  medium  is  placed  in  a  sterile  fermenta- 
tion tube,  and  the  C02  evolved  is  measured  after  a  definite  incuba- 
tion period,  being  compared  with  controls  in  which  no  vitamine  was 
added.  Orange  juice,  yeast  extract,  peptone,  Liebig's  meat  ex- 
tract, honey,  certain  vegetable  extracts  and  milk  were  tried  as  a 
source  of  vitamine,  and  the  results  were  for  the  most  part  the  same 
as  had  been  found  in  animal  experiments.  Certain  discrepant  re- 
sults could  be  explained  by  the  greater  sensitivity  of  the  method. 
Pasteurization  and  sterilization  decrease  the  activity  of  milk.  It 
was  also  interesting  in  this  work  to  note  the  different  behavior  of  the 
two  kinds  of  yeast  as  to  their  dependence  upon  the  vitamine  addi- 
tion, one  requiring  more  than  the  other.  Abderhalden  and  Schau- 


THE   VITAMINES   IN  THE   VEGETABLE   KINGDOM  55 

mann  (I.e.  126)  developed  a  somewhat  similar  method  simultaneously 
with  the  American  investigators. 

There  seems  to  be  no  unanimity  as  to  whether  or  not  yeast 
requires  extraneous  vitamine  for  growth.  Thus,  Fulmer,  Nelson 
and  Sherwood  (129a)  believe  that  yeast  does  not  require  vitamine 
B  for  growth.  These  authors  were  able  to  grow  yeast  on  a 
synthetic  culture  medium  supposedly  free  from  vitamine.  How- 
ever, their  findings  are  not  as  definite  as  their  conclusions.  Nelson, 
Fulmer  and  Cessna  (129b)  have  found  that  if  the  brewer's  yeast 
in  a  synthetic  diet  was  replaced  by  yeast  grown  on  a  synthetic 
medium  in  the  absence  of  vitamines,  the  growth  of  rats  proceeded 
normally,  showing  that  the  above  yeast  synthetized  its  own 
vitamine.  Working  with  pigeons,  Harden  and  Zilva  (129c)  also 
found  that  yeast  synthetizes  vitamine  on  a  synthetic  medium 
but  to  a  lesser  extent  than  in  the  presence  of  vitamine.  Funk  and  . 
Dubin  (129d)  were  unable  to  corroborate  the  findings  of  Fulmer, 
Nelson  and  Sherwood,  using  the  medium  F  described  by  the  latter. 
Poor  yeast  growth  was  obtained  in  the  absence  of  vitamine  while 
a  marked  improvement  was  noted  when  vitamine  was  added.  These 
same  conclusions  were  reached  by  Eddy,  Heft,  Stevenson  and 
Johnson  (129e).  MacDonald  and  McCollum  (129f)  maintain 
also  that  they  can  grow  yeast  on  a  pure  nutrient  medium  in  the 
absence  of  vitamine  B.  However,  they  obtained  better  growth 
with  this  vitamine  added.  This  subject  became  a  matter  of  con- 
troversy between  Ide  (129g)  and  MacDonald  and  McCollum  (129h), 
the  former  maintaining  that  yeast  can  grow  in  two  ways — slow 
growth  without  bios  and  rapid  growth  in  the  presence  of  bios, 
while  the  latter  hold  that  there  is  no  necessity  for  assuming  the 
existence  of  a  specific  substance  promoting  the  growth  of  yeast. 
We  personally  are  in  accord  with  the  view  of  Ide  that  at  least  cer- 
tain yeast  species  do  require  a  specific  substance  for  growth.  A 
number  of  procedures  such  as  improving  the  inorganic  moiety  of 
the  medium  and  the  addition  of  glucose  or  amino  acids  may  give 
slight  increase  in  growth  but  not  to  be  compared  with  that  ob- 
tained on  the  addition  of  vitamine.  Until  we  know  how  many  new 
cells  can  grow  out  of  a  few  broken  down  cells,  this  question,  as  to 
whether  yeast  can  grow  without  vitamine,  must  be  left  open.  From 
the  foregoing,  it  would  seem  that  yeast  cells  can  produce  vita- 
mines,  if  an  original  stimulus  is  provided. 


56  THE   VITAMINES 

BACTERIA 

While  yeast  extract  was  used  in  1904  by  Bertrand  (130)  for  the 
growth  of  B.  xylinum,  it  did  not  occur  to  anyone  that  this  nutritive 
medium  possessed  such  special  characteristics.  Most  bacteria,  how- 
ever, thrive  well  on  the  commonly  used  media.  With  some  organisms 
on  the  contrary,  difficulties  arise  and  consequently  the  method  of 
Noguchi  (131),  recommending  the  addition  of  a  small  amount  of 
testicular  substance  for  the  cultivation  of  spirochetes,  denotes  impor- 
tant progress.  What  was  responsible  for  the  stimulating  effect  of 
this  tissue?  The  question  remained  unanswered  till  the  war 
came  and  the  bacteriological  laboratories  were  stripped  of  their 
customary  sources  of  supply  of  nutritive  media.  It  became  neces- 
sary then,  above  all  things,  to  replace  Witte's  peptone  and  nutrose. 
Investigators  found  this  difficult,  till  systematic  work  on  this  subject 
was  undertaken.  On  the  whole,  it  may  be  said  that  most  of  the 
nutritive  media  used  before  the  war  very  likely  owed  their  favorable 
influence  on  the  growth  of  bacteria  to  a  small  amount  of  vitamine 
present  therein.  This  conclusion  was  not  apparent  immediately, 
since  many  investigators  thought  that  bacteria  could  grow  on  a  pure 
synthetic  medium.  For  instance,  Vedder  (132)  cultivated  the 
meningococcus  on  starch  prepared  from  cornmeal.  Doryland  (133) 
reported  success  with  pure  synthetic  media,  which  evidently  con- 
tained no  vitamine.  These  experiments  may  be  explained  in  that 
the  inoculation  was  perhaps  so  great,  that,  similar  to  yeast,  growth 
cook  place.  Pieper,  Humphrey  and  Acree  (134)  reported  great 
success  with  synthetic  media,  while  Lockeman  (135)  considered  that 
the  factors  necessary  for  the  growth  of  the  tubercle  bacillus  were 
asparagine,  as  a  source  of  nitrogen,  glycerol  and  inorganic  salts, 
phosphorus,  potassium  and  magnesium.  Nevertheless,  for  other 
microorganisms,  other  requirements  appeared  to  be  necessary,  and 
Bainbridge  (136)  believed  that  certain  microorganisms  could  not 
grow  with  protein  as  the  only  source  of  nitrogen.  This  view  was 
also  maintained  by  Sperry  and  Rettger  (137).  Robinson  and 
Rettger  (138)  compared,  in  this  connection,  a  protein  obtained  by 
ferment  action  (which  he  called  "Opsin"),  with  the  products  of 
protein  acid  hydrolysis.  Decolorization  of  the  opsin  by  animal 
charcoal  decreased  the  value  of  that  nutritive  medium.  Of  the 
proteins  tested,  the  products  of  casein  acid  hydrolysis  gave  the  best 
results,  but  not  so  good  as  opsin.  Extracts  of  beef  worked  very  welL 


THE   VITAMINES   IN   THE   VEGETABLE   KINGDOM  57 

It  was  of  greatest  importance  to  this  entire  subject  that  the 
presence  of  vitamines  in  typhus  bacilli  was  detected  by  Pacini  and 
Russel  (139).  They  proceeded  from  the  observation  that  in  the 
typhus  convalescence  period  there  is  often  noticeable  quite  an  increase 
in  growth.  The  bacilli  were  grown  on  the  usual  vitamine-free 
Uschinsky  nutritive  medium,  and  consequently  they  must  have  been 
able  to  synthesize  the  necessary  vitamine.  Both  the  remaining 
medium  and  an  acid-alcoholic  extract  of  the  bacteria  were  admin- 
istered to  rats  on  an  artificial  diet,  whereupon  normal  growth  took 
place.  Since  the  presence  of  vitamines  in  bacteria  was  demonstrated 
in  this  way,  it  was  plausible  to  assume  that  for  certain  types  of 
bacteria,  a  vitamine-containing  medium  would  be  of  value.  Such  a 
preparation  was  introduced  by  Mackenzie  Wallis  (140) ;  it  consisted 
of  casein,  peanut  flour  and  soda,  and  served  as  a  substitute  for  nutrose 
in  the  composition  of  Conradi-Drygalski  medium  (for  the  typhus 
coli  group) .  The  bacteria  grew  well  on  the  medium  and  the  success 
was  attributed  to  the  presence  of  vitamine  in  the  globulin  fraction 
of  the  flour.  A  similar  preparation,  introduced  by  Huntoon  (141) 
was  made  from  meat  and  eggs.  In  the  description  of  this  preparation 
it  was  stipulated  that  all  filtration,  whether  through  filter  paper  or 
cotton,  must  be  avoided.  A  yeast  extract  as  culture  medium  was 
also  described  by  Ayers  and  Rupp  (142). 

Meningococcus.  The  conditions  for  the  growth  of  this  micro- 
organism were  systematically  studied  by  Miss  Lloyd  (143).  She 
found  that  growth  was  impossible  without  vitamines.  Although 
amino  acids  were  the  chief  components  of  the  nutritive  medium,  it 
could  not  be  utilized  except  when  a  voluminous  inoculation  had 
been  made.  In  this  way  it  was  possible  to  note  a  difference  in  the 
behavior  of  different  laboratory  stock  toward  vitamines — older 
laboratory  stock  needed  no  special  vitamine  addition  when  a  sufficient 
amount  of  amino  acids  was  available,  while  newly  isolated  bacteria 
could  not  be  cultivated  without  vitamine.  Between  the  necessary 
amounts  of  vitamine  and  amino  acids,  some  relationship  is  apparent, 
and  therefore  it  is  held  that  the  role  of  the  vitamines  in  the  nutrition 
of  the  meningococcus  is  traceable  to  the  acceleration  of  the  reaction 
velocity  of  the  proteolytic  processes.  A  further  function  is  attributed 
to  the  vitamines  in  that  the  assimilation  of  iron,  calcium,  phosphorus 
and  iodine  is  facilitated.  The  vitamine  in  question  was  soluble  in 
water  and  alcohol  and  somewhat  thermo-stable ;  it  could  stand  heat- 


58  THE   VITAMINES 

ing  for  45  minutes  at  120°C.  This  vitamine  could  be  easily  adsorbed 
by  filter  paper  but  not  by  glass  wool.  It  was  shown  to  be  present 
in  blood  and  in  milk  (and  to  a  smaller  extent  in  serum).  On  this 
basis,  a  blood  agar  was  prepared  (144)  which,  in  spite  of  the  coagula- 
tion of  its  protein  by  heat,  still  contained  vitamine,  but  could  not  be 
subjected  to  filtration.  The  precautions  to  be  observed  as  regards 
filtration  pointed  to  the  probability  that  they  were  dealing  with  two 
vitamines,  both  of  which  were  necessary  and  one  of  which  was  either 
slightly  soluble  in  water  or  not  at  all.  The  latter  could  be  adsorbed 
by  protein,  and  of  special  interest,  is  the  fact  (145)  that  the  meningo- 
coccus,  after  from  one  to  ten  re-inoculations,  does  not  need  any 
more  vitamine  additions,  and  this  is  apparently  the  reason  why  the 
importance  of  vitamines  for  bacteria  was  not  recognized  sooner.  At 
the  same  time,  somewhat  similar  experiments  were  carried  out  by 
Gordon  and  Hine  (146),  and  also  by  Flack  (147).  These  investi- 
gators were  able  to  show  that  serum  albumin,  hemoglobin,  fibrinogen, 
legumin  and  human  blood  accelerated  the  growth  of  the  meningo- 
coccus  on  common  agar,  while  the  addition  of  glucose  was  without 
any  effect.  Among  others,  they  recommended  trypagar,  prepared 
from  pea  flour  and  digested  ox  heart.  The  best  results  were  obtained 
with  an  agar  containing  bouillon  and  an  extract  of  cooked  pea  flour, 
or  the  fractions  obtained  after  removal  of  starch  and  dextrin  by 
alcohol.  Extracts  of  raw  and  cooked  wheat  embryo  showed  a 
similar  ability  to  promote  growth  and  to  prolong  viability;  these 
authors  believed  that  pea  flour  contained  a  vitamine  acting  on  growth, 
while  that  from  wheat  influenced  the  viability.  Shearer  (148)  also- 
investigated  the  influence  of  some  extracts.  He  found  the  active 
substance  in  nasal  mucous  and  believed  that  it  acted  not  as  a  usual 
dietary  constituent,  but  as  an  activator.  The  substance  was  soluble 
in  water,  less  soluble  in  alcohol  and  not  at  all  in  ether;  it  was 
unaffected  by  heating  with  strong  hydrochloric  acid  for  12  hours. 
Besides  the  meningococcus,  it  was  active  for  pneumococcus,  B. 
typhosus,  B.  coli  communis  and  other  pathogenic  bacteria.  In  the 
continuation  of  his  work,  Shearer  (149),  noted  that  the  cerebrospinal 
fluid  contained  a  substance  which  is  still  more  active  than  blood  or 
nasal  mucus.  This  peculiarity  explained  the  rapid  growth  of-  the 
meningococcus  in  the  brain  tissues  and  in  the  membranes  and 
cavities  of  the  central  nervous  system.  In  this  case  too,  the  action, 
was  observed  not  upon  the  meningococcus  only,  but  also  upon  other 


THE    VITAMINES   IN   THE    VEGETABLE    KINGDOM  59 

pathogenic  bacteria,  like  pneumococcus,.B.  typhosus,B.  coli  communis, 
certain  intestinal  streptococci  and  throat  bacteria.  The  proof  that 
the  vitamine  was  actually  necessary  was  not  available,  acceleration 
in  growth  only  being  demonstrated.  Eberson  (150)  likewise  found 
that  a  yeast  extract  could  increase  the  viability  of  a  meningococcus 
culture  for  more  than  a  month.  The  medium  consisted  of  agar, 
peptone  and  potassium  phosphate.  The  yeast  was  first  extracted  for 
two  hours  at  not  more  than  100°C.,  and  then  sterilized  for  a  half 
hour  at  one  atmosphere. 

Gonococcus.  Cole  and  Lloyd  (151)  carried  out  vitamine  experi- 
ments on  gonococcus.  Tryptamine  blood  (tryptic  digestion  product 
of  casein)  was  used  for  cultivation.  The  gonococci  cultures  required 
a  suitable  ion  concentration,  a  considerable  amount  of  free  amino 
acids  and  two  kinds  of  vit amines  to  permit  of  favorable  development. 
One  of  these,  easily  adsorbable,  is  found  in  blood  corpuscles  and 
possesses  the  ability  to  promote  growth;  the  other  occurs  in  tissues, 
is  relatively  little  adsorbable,  and  has  the  peculiarity  of  bringing 
about  a  secondary  plentiful  growth.  The  method  of  demonstration 
of  both  vitamines  is  given  in  great  detail.  They  are  precipitated  by 
protein,  but  not  in  the  presence  of  agar.  Filtration  through  paper 
removes  the  first  vitamine  completely  and  we  have  a  good  basis  for 
the  idea  that  the  substance  from  the  blood  corpuscles  belongs  to  the 
type  of  A-vitamine  (antirachitic  vitamine)  which  is  found  naturally 
associated  with  fats;  or  it  may  also  belong  to  a  vitamine  type  that 
is  easily  adsorbed  by  protein.  On  the  other  hand,  the  vitamine  of  the 
tissues  belongs  perhaps  to  the  B-vitamine  type.  Blood  seems  to 
contain  both  substances  although,  in  sheep  serum,  the  first  one 
appears  to  be  lacking.  Morini  (152)  grew  the  gonococcus  on  gelatin 
and  beer  yeast. 

The  observations  of  Douglas,  Fleming  and  Colebrook  (153)  suffice 
to  give  us  some  idea  of  the  conditions  under  which  anaerobic  bacteria 
are  best  cultivated.  Special  measures  for  the  exclusion  of  air  are 
shown  not  to  be  necessary,  if  the  bacteria  are  given  the  opportunity 
to  provide  themselves  with  local  anaerobic  conditions  by  the  addition 
of  some  porous  material  such  as  asbestos.  It  is  very  important, 
however,  to  add  to  the  bouillon  culture,  potatoes,  carrots,  cabbages, 
and  grapes,  or  better  still,  bran.  An  alcoholic  extract  of  bran  is 
effective  only  when  large  amounts  are  used. 


60  THE   VITAMINES 

Tubercle  bacilli  were  cultivated  by  Sazerac  (153a)  and  by  Masucci 
(153b)  on  digested  casein  (aminoids). 

In  regard  to  hemophilic  bacteria,  there  is  the  work  of  Davis  (154). 
For  this  type,  two  factors  in  particular  are  of  great  importance.  One 
is  hemoglobin,  and  the  other  is  a  substance  which  is  found  in  other 
strange  bacteria,  as  well  as  in  a  variety  of  fresh  plant  and  animal 
tissues.  This  second  substance  is  found  in  B.  influenza  when  both 
strains  are  cultivated  together.  The  similarity  of  the  factors  neces- 
sary for  the  growth  of  rats  with  those  necessary  for  the  successful 
growth  of  hemophilic  bacteria  has  been  emphasized  by  Davis.  He 
explained  it  by  the  fact  that  hemoglobin  serves  as  the  source  of  iron, 
while  the  second  substance  facilitates  the  assimilation  of  this  element. 
In  a  later  publication,  Davis  (155)  found  that  the  growth-promoting 
influence  of  hemoglobin,  white  and  brown  rice,  pure  wheat  flour  and 
wheat  bran,  was  shown  by  no  other  kinds  of  bacteria  except  by  the 
hemophylic.  Germinated  rice  or  wheat  kernels  exhibit  a  greater 
influence  than  ungerminated.  In  this  case  also,  the  active  substance 
was  adsorbable  by  filter  paper  and  was  quite  thermo-stable,  having 
resisted  a  temperature  of  100°C.  for  one  to  two  hours. 

The  favorable  influence  of  cooked  meat  on  bacterial  growth  was 
observed  by  Wolf  and  Harris  (156),  on  the  B.  histolyticus,  and  by 
Wolf  (157)  on  the  vibrion  septicus,  B.  sporogenes,  B.  welchii  and  B. 
proteus  (158).  Boyer  (159)  recommended  a  hydrochloric  acid  extract 
of  finely  divided  bones  for  the  cultivation  of  streptococci,  and  Otabe 
(160)  suggested  a  wheat  extract  instead  of  meat  for  staphylococci. 
We  can  see  from  this  short  list  that  the  addition  to  the  medium  of  a 
natural  product  as  a  source  of  vitamine  was  necessary.  For  the 
cultivation  of  B.  coli,  Dienert  and  Guillerd  (161)  prepared  a  medium 
as  follows:  500  grams  of  pressed  yeast  were  heated  to  50°C.,  the 
optimum  temperature  for  the  action  of  endotryptase.  In  less  than 
24  hours  the  autolysis  was  finished,  the  yeast  having  become  liquid, 
yielding  397  cc.  of  filtrate  which  was  then  diluted  to  two  liters.  The 
extract  was  sterilized  for  a  half  hour  at  100°C.,  and  finally  diluted  to 
7500  cc.  This  preparation  was  used  advantageously  instead  of 
peptone. 

Bunker  (162)  and  particularly  L.  David  and  Ferry  (163)  have 
made  an  exact  study  of  the  conditions  that  are  necessary  in  order  to 
obtain  a  very  active  toxin  from  cultures  fo  B.  diphtherias.  It  was 


THE   VITAMINES   IN   THE   VEGETABLE   KINGDOM 


61 


shown  by  these  investigators  that  a  medium  of  amino  acids  and  salts 
did  not  suffice  and  that  an  addition  of  creatine,  creatinine,  xanthine 
and  hypoxanthine  had  no  effect.  Only  when  a  0.5  per  cent  of 
bouillon  was  added,  did  it  result  in  good  growth.  For  the  prepara- 
tion of  toxins,  the  amount  of  bouillon  present  must  be  increased  to  10 
per  cent.  Addition  of  peptone,  on  the  contrary  resulted  in  a  slight 
growth  and  toxin  formation.  Toxin  is  not  supposed  to  be  a  synthetic 
product  of  bacterial  cells;  but  a  product  of  metabolism  which  makes 
its  appearance  only  in  the  presence  of  certain  amino  acids  and 
vitamine. 

Agulhon  and  Legroux  (164)  investigated  the  influence  of  vitamine 
on  B.  influenzae  (B.  Pfeiffer).  The  growth-accelerating  influence  of 
blood,  serum  and  ascitic  fluid,  could  not  have  been  due  to  protein 
addition  but  to  the  influence  of  vitamine.  The  vitamine  extract  of 
defibrinated  blood  was  obtained  in  two  ways.  By  the  first,  blood 
was  precipitated  by  about  four  volumes  of  absolute  alcohol;  the 
precipitate  was  extracted  with  the  same  volume  of  water,  centrifuged 
and  filtered  through  a  Chamberlain  filter.  By  the  second  procedure, 
blood  was  extracted  with  sodium  chloride  solution,  and  the  extract 
heated  to  80°C.  for  15  minutes.  From  5  to  10  per  cent  of  these 
extracts,  added  to  the  usual  medium,  showed  a  great  influence  on 
the  growth  of  B.  Pfeiffer.  Even  a  one  per  cent  solution  showed  a 
definite  effect.  It  might  well  be  conceived  that  the  vitamines  in 
the  cells  are  present  in  combination  and  that  alcohol  or  heating  sets 
them  free.  They  seemed  to  be  insoluble  in  alcohol,  for  a  cold 
alcoholic  extract  was  inactive,  while  one  prepared  at  80°C.  was  only 
slightly  active.  Acetone  extracts,  prepared  either  hot  or  cold,  were 
inactive.  Warming  to  80°C.  in  the  presence  of  alcohol  or  acetone, 
with  subsequent  drying  at  a  lower  temperature  did  not  bring  about 
decomposition,  since  extraction  of  this  powder  with  the  water  yielded 
an  active  substance.  The  watery  vitamine  extract  partially  lost  its 
activity  after  heating  to  90°C.  for  15  minutes.  It  appears  to  be  more 
stable  when  the  heating  is  done  in  the  presence  of  gelose,  although  in 
this  case  too,  it  is  partially  or  completely  inactivated,  depending  upon 
whether  the  temperature  is  raised  only  to  100°or  to  120°C.  When 
a  totally  defibrinated  blood  was  used,  it  was  observed  that  the  vita- 
mines  were  found  in  the  corpuscles,  since  the  watery  extract  of  the 
serum  gave  completely  negative  results,  while  the  washed  blood 
corpuscles  gave  positive  results.  Hemoglobin  and  blood  corpuscle 


62  THE    VITAMINES 

stroma  were  negative.  Cold  extraction  of  blood  corpuscles  for  24 
hours  with  a  physiological  salt  solution  gave  only  a  weakly  active 
extract.  If  hemolysis  is  induced  with  distilled  water  before  the 
extraction,  then  the  vitamines  are  found  in  solution.  In  this  case, 
contrary  to  what  has  been  noted  previously  in  this  chapter,  the 
extracts  may  be  filtered  through  a  Chamberlain  filter  or  through 
paper  without  loss  of  activity;  this  characteristic  permits  of  steriliza- 
tion without  the  use  of  heat.  Incidentally,  the  action  of  vitamines 
has  been  erroneously  ascribed  to  their  peculiar  physical  condition. 
Legroux  and  Mesnard  (165)  extended  their  experiments  to  include 
extracts  of  kidney,  liver  and  heart.  Davis  (165a)  has  pointed  out 
the  necessity  of  two  substances  for  the  growth  of  B.  influenzae,  one 
derived  from  the  blood  pigment  and  one  of  a  vitamine  nature.  The 
same  conclusions  were  reached  by  Rivers  and  Poole  (165b),  by 
Thjotta  (165c)  and  by  Thjotta  and  Avery  (165d).  Thjotta  and 
Avery  differentiate  between  a  thermo-stable  substance  contained 
in  blood,  and  called  "A,"  and  a  thermo-labile  substance  "V"  (vita- 
mine)  which  is  found  in  blood,  yeast  and  vegetable  extracts.  Crys- 
talline hemoglobin  contains  only  the  substance  "  A." 

Finally,  there  are  the  vitamine  studies  of  Kligler  (166)  on  bacteria. 
At  first  he  used  autolyzed  yeast  and  found  that  pneumococci  and 
meningococci  do  not  grow  so  well  on  it  as  on  bouillon.  In  a  later 
work,  he  (167)  investigated  the  influence  of  ox  heart,  goat  blood, 
rabbit  and  cat  tissue  and  human  excreta  on  gonococcus,  meningo- 
coccus,  pneumococcus,  Streptococcus  hemolyticus,  B.  diphtheriae,  B. 
pertussis  and  B.  influenzae.  All  of  the  investigated  extracts  exerted 
a  favorable  action  in  exact  proportion  to  the  amount  added.  The 
spleen,  liver  and  kidneys  were  rich  in  vitamines,  while  muscle 
tissue  was  only  slightly  active,  and  ether  extracts  were  inactive. 
Mueller  (167a)  found  that  streptococcus  needed  for  growth  a  sub- 
stance which  he  regarded  as  a  new  amino  acid.  The  streptococcus 
was  cultivated  on  beef-heart  extracts.  If  this  medium  was  de- 
colorized with  "norit"  it  was  unsuitable  for  growth,  but  permitted 
of  growth  upon  the  addition  of  peptone  or  certain  protein  hydrol- 
yzates.  The  growth  substance  was  precipitated  from  these  hydrol- 
yzates  with  mercuric  sulphate,  but  it  was  not  identical  with 
tryptophane,  tyrosine,  cystine  or  histidine.  In  a  subsequent 
publication  the  same  author  (167b)  accepted  the  necessity  of  three 
substances  for  the  growth  of  Streptococcus  hemolyticus — one  which 


THE    VITAMINES    IN    THE    VEGETABLE    KINGDOM 


63 


remains  in  the  decolorized  beef-heart  infusion  and  two  others  which 
were  found  in  the  hydrolyzate  of  the  commercial  casein  preparation 
"aminoids."  Of  the  latter  two,  the  substance  "x"  was  detected 
in  the  silver  sulphate  precipitate  and  the  substance  "y"  in  the  fil- 
trate. In  collaboration  with  Freedman  (167c),  we  were  able  to 
show  that  not  all  pure  proteins  (hydrolyzed)  fulfill  the  nutritional 
requirements  of  Streptococcus  hemolyticus.  As  a  rule,  purification 
of  these  proteins  diminished  their  nutritive  value  for  these  bac- 
teria. Other  proteins  obtained  from  sources  rich  in  vitamines 
are  with  great  difficulty,  or  not  all,  freed  from  this  growth  substance. 
On  the  other  hand,  this  growth  substance  can  be  adsorbed  in  a  way 
similar  to  vitamines,  since  fuller's  earth  and  norit  remove  it  from 
autolyzed  yeast,  hydrolyzed  casein  and  a  neutral  solution  of  sodium 
oaseinate.  This  phenomenon  will  permit  of  further  investigation. 
The  absence  of  the  growth  substance  from  a  number  of  purified 
proteins  of  various  sources  renders  it  unlikely  that  the  above  growth 
factor  is  an  integral  part  of  the  protein  molecule.  It  is  more  than 
likely  that  the  growth  substance  is  similar  to  that  necessary  for 
yeast — and  found  in  the  same  starting  material — the  only  dis- 
crepancy being  that  those  hydrolyzed  proteins  which  act  on  strepto- 
coccus do  not  act  upon  yeast.  However,  this  might  easily  be  a 
matter  of  inhibition  due  to  the  various  protein  cleavage  products. 
In  this  connection,  Norris  (167d)  found  that  casein  was  a  better 
medium  than  brewer 's  yeast  for  culture. 

The  many  points  brought  out  in  this  small  chapter,  touching  in 
particular  upon  the  physical  characteristics  of  the  substances  which 
act  on  bacteria,  form,  for  the  time  being,  the  sole  criterion  for  their 
classification  in  the  group  of  vitamines.  By  comparing  the  char- 
acteristics here  set  forth  with  the  description  of  the  vitamines  to 
follow  later,  the  reader  will  have  the  opportunity  of  making  a  decision 
for  himself  as  to  which  group  these  substances  should  be  assigned. 
Looking  upon  this  chapter  as  a  whole,  we  may  conclude  that  at  least 
some  types  of  bacteria  respond  to  the  addition  of  vitamine  with 
accelerated  growth,  showing  possibly  that  a  small  initial  quantity  of 
vitamine  suffices  to  render  the  bacteria  independent  of  any  further 
vitamine  additions  for  a  long  time.  As  we  shall  see  in  the  chapter 
on  the  growth  of  higher  plants,  bacteria  seem  to  exist  entirely  inde- 
pendent of  any  vitamine  supply.  According  to  our  present  knowl- 
edge, these  are  the  bacteria  which  supply  the  whole  organized  world 


64  THE   VITAMINES 

with  the  vitamines,  or  with  the  substances  from  which  they  are 
formed. 

All  of  the  above  mentioned  observations  conform  preferably  with 
the  conception  of  two  or  three  different  vitamines. 

FUNGI 

Because  of  the  lesser  practical  significance  as  compared  with 
bacteria,  the  question  as  to  the  vitamine  requirements  of  fungi 
received  little  attention.  Lutz  (168)  made  the  observation  that  fungi 
often  show  an  interruption  in  growth  when  the  nutritive  medium  is 
exhausted.  Still,  this  observation  could  be  attributed  to  a  general 
exhaustion  of  nutritive  constituents  in  the  medium.  According  to  a 
paper  by  Currie  (169),  Aspergillus  niger  needs  no  extra  addition  in 
order  to  grow,  and  yet  he  states  that  an  extract  of  old  mycellium,  or 
spore  bearing  portions  of  the  same  fungus,  brings  about  quite  a 
definite  acceleration  in  the  growth  of  a  new  culture  of  the  same 
fungus. 

The  only  work  affording  an  insight  into  the  vitamine  requirements 
of  parasitic  and  saprophytic  fungi  is  the  recent  investigation  of 
Willaman  (170) .  He  worked  with  Sclerotinia  cinerea  [(Bon)  Schroter],. 
which  he  sought  to  cultivate  on  a  solution  of  purified  salts,  cane 
sugar  and  nitrates.  This  experiment,  and  others  in  which  there 
was  also  an  addition  of  asparagine  and  amino  acids,  turned  out 
negative.  The  results  were  different  when  fruit  juices,  yeast  or 
extracts  of  wheat  germ  were  used.  The  method  of  procedure  was  so 
chosen  that  the  surface  of  the  mycellium  was  measured  in  square 
centimeters,  and  the  number  of  spores  noted.  The  juices  from 
peaches,  plums  and  apples  showed  the  greatest  activity.  Besides- 
these,  a  great  number  of  natural  products  were  tested  and  found 
active,  at  least  as  regards  the  growth  of  this  fungus.  However,  not 
all  of  these  extracts  were  equally  active  on  the  sporulation.  For 
this  purpose,  extracts  had  to  be  prepared  from  parts  of  plants  which 
displayed  a  very  active  metabolism.  The  following  could  be  classed 
as  such:  pollen,  sporophores  of  the  fungi  and  the  sclerotinia  itself, 
terminal  butts  of  the  Phaseolus  and  sporulated  mycellium  of  Asper- 
gillus. This  observation  indicates,  perhaps,  the  importance  of  the 
two  types  of  vitamine,  although  Willaman  is  inclined  to  assume,  in 
the  meantime,  the  existence  of  a  single  vitamine,  varying  in  its  effect 
according  to  its  source  of  origin.  Nevertheless,  one  must  concede 


THE    VITAMINES   IN   THE   VEGETABLE    KINGDOM  65 

that  in  the  course  of  this  work,  the  firmly  established  physical  and 
chemical  characteristics  of  the  active  substances  are  best  in  accord 
with  the  conception  of  a  second  vitamine.1  Working  with  Oidium 
lactis,  Linossier  (170a)  found  that  on  addition  of  vitamine,  the 
activation  took  place  only  in  the  first  few  days. 

HIGHER   PLANTS 

In  the  first  edition  of  this  book,  in  anticipation  of  a  possible 
analogy  to  animal  growth,  we  wrote  a  small  chapter  on  plant 
growth  without  being  able  to  find  any  supporting  experimental 
evidence;  still,  it  was  not  necessary  to  wait  for  it  a  long  time.  In 
1909,  Clinton  (171)  made  the  observation  that  plant  extracts  have 
the  power  to  stimulate  the  reproduction  of  certain  plants.  Haber- 
landt  (172)  found  that  parts  of  potatoes  containing  the  morphological 
structure,  "Leptom,"  showed  cell  division  when  placed  in  a  nutritive 
solution.  The  embryonic  structure  only  was  able  to  undergo  cell 
division  without  the  presence  of  leptom.  This  author  suspected  the 
presence  of  a  special  hormone  in  leptom  corresponding  to  the  internal 
secretions  of  animals. 

Bottomley  (173)  was  the  first  investigator  to  take  up  this  problem 
seriously.  In  1912,  he  occupied  himself  with  the  problem  of  the 
ability  of  certain  bacteria  to  transform  the  humic  acids  of  peat  into 
water-soluble  combinations.  He  demonstrated  at  that  time  that 
such  peat  contains  all  of  the  chemical  combinations  necessary  for  the 
growth  of  young  tomato  plants,  buckwheat,  radishes,  and  barley. 
In  the  further  development  of  this  question,  which  appeared  at 
about  the  same  time  as  our  earlier  vitamine  work,  Bottomley  (174) 
extracted  a  substance  from  bacterized  peat  with  water  or  alcohol. 
The  addition  of  phosphotungstic  acid  to  this  alcoholic  extract  yielded 
a  precipitate  which,  upon  decomposition,  acted  favorably  on  plant 
growth.  The  solution  thus  obtained  was  precipitated  with  silver 
nitrate,  and  in  the  end  the  growth  substance  was  found  present  in 
the  same  fraction  as  the  vitamine.  All  active  fractions  were  tested 
for  their  influence  on  the  growth  of  young  plants,  and  then,  in  order 
to  set  forth  the  results  more  clearly,  the  cotyledons,  which  contain 

1  Willaman  defines  the  vitamines  as  a  class  of  substances  whose  presence 
is  necessary  for  normal  metabolism,  but  which  do  not  contribute  to  the 
requirements  of  the  organism  as  regards  inorganic  constituents,  nitrogenous 
substances,  and  energy  producing  food  constituents. 


66 


THE   VITAMINES 


reserve  vitamine,  were  removed  after  germination  without  harm  to 
the  plant.  The  results  were  very  significant  and  showed  that  the 
young  plants,  in  a  stage  in  which  they  could  not  live  by  their  own 
process  of  metabolism,  are  dependent  upon  the  stock  of  vitamine 
which  is  stored  up  in  the  seeds.  In  mature  plants,  the  vitamines 
are  built  up  through  symbiosis  with  bacteria,  which  prepare  it 
synthetically  from  the  organic  constituents  of  the  soil  and  from 
8 


O) 

.£       6 
•+-> 

_0) 

i 


A 


B 


o  10  ao  Jo  40  50 

Time  in  days 
FIG.  4.  PLANT  GROWTH 
A,  with  vitamine  addition;  B,  without  vitamine  addition   (Bottomley) 

fertilizers,  and  carry  it  to  the  plants  through  the  roots.  In  the 
investigations  of  Bottomley,  the  control  plants  (in  the  nutritive 
solution  above)  underwent  a  decrease  in  weight  of  10.9  per  cent  in  50 
days,  while  with  the  addition  of  vitamine  there  was  an  increase  in 
weight  of  59  per  cent.  The  curves  appended  illustrate  what  has  just 
been  stated.  We  see  here  the  influence  of  the  silver  nitrate  fraction 


THE    VITAMINES   IN   THE    VEGETABLE    KINGDOM  67 

added  to  the  nutritive  solution  in  a  dilution  of  0.35  in  1,000,000. 
Bottomley's  results  were  soon  confirmed  by  others  (175).  The  sub- 
stances influencing  plant  growth  were  called  "auximones"  by  Bot- 
tomley,  who  also  described  a  method  for  their  detection  (176).  This 
method  possesses  certain  advantages  as  against  the  use  of  higher 
plants,  and  consists  in  an  extraction  of  fermented  peat,  and  precipita- 
tion of  the  resulting  extract  with  phosphotungstic  acid.  If  this 
extract  is  added  to  a  culture  of  nitrifying  bacteria  and  the  whole 
placed  in  an  incubator  at  26°C.,  a  scum  is  formed  on  the  surface 
after  24  to  36  hours,  and  no  nitrates  are  found  in  the  solution.  On 
the  contrary,  without  the  addition  of  vitamine,  the  scum  does  not 
appear  and  the  nitrification  proceeds  rapidly  instead.  This  method 
seems  to  be  specific  for  the  vitamines,  since  no  such  effect  has  been 
recorded  after  the  addition  of  cane  sugar,  maltose,  asparagine, 
peptone,  leucine,  tyrosine  and  hordeine.  The  microorganism  isolated 
from  this  scum,  similar  to  the  nitrifying  and  sulphur-  and  iron- 
assimilating  bacteria,  may  grow  without  the  addition  of  organic 
carbon  combinations.  Originally,  Bottomley  was  of  the  opinion  that 
the  auximones  differed  from  the  vitamines  in  that  the  former  were 
heat  resistant.  We  believe,  however,  that  a  successful  classification 
cannot  be  built  up  on  such  differentiations.  Chittenden  (177) 
repeated  these  flower-pot  experiments  in  which  the  ratio  of  the 
bacterized  peat  to  the  required  amount  of  soil  was  very  high,  and 
therefore  was  also  able  to  produce  an  acceleration  of  growth.  When 
this  method  of  experimentation  was  applied  to  greatly  extended 
soil  experiments,  the  results  were  not  so  significant;  the  greatest 
success  was  obtained  when  the  rainfall  was  large.  Bottomley  (178) 
himself  repeated  his  first  experiment  more  carefully.  For  this 
purpose  he  used  a  type  of  lentil  (Lemna  minor)  that  could  live  in 
water;  this  was  cultivated  on  Detmer's  nutritive  solution,  of  which 
the  only  source  of  nitrogen  was  potassium  nitrate.  This  plant  could 
not  live  only  upon  inorganic  constituents;  it  gradually  became 
weaker,  and  presented  an  abnormal  appearance.  Bottomley  then 
added  to  the  nutritive  solution  various  extracts  prepared  from 
fermented  peat.  From  the  results  recorded  in  the  table  below,  only 
one  conclusion  may  be  drawn — that  the  influence  of  these  extracts 
was  due  not  only  to  the  presence  of  certain  necessary  building  stones 
therein,  but  also  to  the  presence  of  vitamines.  These  experiments 
were  then  repeated  by  Bottomley  (179)  with  other  water-plants  like 
Selvinia  natans,  Azolla  filiculoides  and  Limnobium  stoloniferum, 
with  the  same  results. 


68 


THE   VITAMINES 


The  extracts  of  the  fermented  peat  were  examined  chemically  by 
Rosenheim  (ISO).2  To  give  the  reader  some  conception  of  the 
quantities  of  the  active  substances  used,  we  shall  go  into  these  experi- 
ments more  in  detail:  15  grams  of  bacterized  peat  were  extracted 
with  2  liters  of  water  and  500  cc.  of  this  brown  solution  were  diluted 
to  1250  cc.  This  extract  contained  0.045  per  cent  of  dry  substance, 
0.034  per  cent  of  organic  and  0-.011  per  cent  of  inorganic  material, 
and  0.003  per  cent  of  nitrogen.  The  plants  investigated  (Primula 
malacoides)  received  in  one  dose  0.18  grams  of  peat,  which  contained 
20  mgm.  of  organic  material  and  1.9  mgm.  of  nitrogen.  The  plants 
so  treated  (figure  5)  grew  higher,  showed  leaves  of  a  deep  green  color, 
and  were  also  generally  larger  than  the  controls,  which  is  especially 
apparent  in  the  illustration.  The  substance  which  could  bring  about 


« 

VELOCITY  OF  WEIGHT 
DOUBLING 

UNITS  OF  TIME  NECES- 
SARY FOR  DOUBLING 
OF  WEIGHT 

Number 
of  plants 

Original 
weight 

Number 
of  plants 

Original 
weight 

Controls                                

1.0 
2.05 
1.91 
1.63 

1.24 

1.0 
3.09 
2.71 
2.01 

1.48 

100 
48 
52 
61 

SO 

100 

32 
37 
50 

67 

Watery  extract  of  bacterized  peat..  .  . 
The  same  without  humus  acid  

Alcoholic  extract  of  bacterized  peat.  .  . 
Phosphotungstic  acid  precipitate  pre- 
pared from  the  last  fraction  

such  a  great  change  must  belong  to  the  type  of  vitamines,  according 
to  Rosenheim.  From  the  active  extract,  it  was  possible  to  obtain 
with  phosphotungstic  acid  a  precipitate  which  was  only  partly 
soluble  in  acetone  (like  the  vitamines,  as  we  shall  see  later) .  Rosen- 
heim hoped  to  extend  his  experiments  to  include  yeast  and  milk. 

Bottomley  (181)  investigated  the  nitrogenous  constituents  of  the 
peat,  and  although  there  was  no  nucleic  acid  present,  he  did  find 
their  decomposition  products,  an  uracil-adenine-dinucleotidc  and  the 
components  of  guanine-cytosine-dinucleotide. 

Appleman  (182)  believed  that  potato  tubers  contained  substances 
which  were  necessary  for  the  growth  of  the  shoots.  It  is  therefore 
of  importance  in  planting  potatoes,  that  not  too  small  a  piece  of  the 


2  In  this  paper,  Rosenheim  claims  priority  as  to  the  findings  on  the  impor- 
tance of  vitamines  for  plants. 


THE   VITAMINES   IN   THE   VEGETABLE   KINGDOM 


69 


tuber  be  used.  He  thought  that  the  disease  of  potatoes  common  in 
the  United  States,  and  known  as  "spindling  sprout  disease,"  was  due 
to  lack  of  these  vitamine-like  substances. 

Bottomley  (183)  also  investigated  the  influence  of  crude  nucleic 
acid  derivatives  upon  the  growth  of  Lemna  minor.     The  action,  in 


FIG.  5.  GROWTH  OF  PRIMULA  AFTER  VITAMINE  ADDITION  (ROSENHEIM) 

this  instance,  was  similar  to  the  activity  of  a  sterilized  suspension  of 
Azotobacter  chroococcum.  The  combined  activity  of  the  two  products 
the  suspension  and  the  nucleotides  were  complementary  to  each 
other.  Purified  adenine-uracil  fractions  showed  no  activity.3  In 


3  Papers' published  by  Jones  (184)  and  also  by  Stead  (185)  may  be  looked 
upon  as  reviews. 


70  THE   VITAMINES 

Bottomley's  laboratory,  Mockeridge  (186)  studied  the  effect  of 
extracts  of  bacterized  peat  upon  nitrifying  bacteria  of  the  soil  (Bacil- 
lus radicicola  and  Azotobacter  chroococcum) .  The  results  are  apparent 
from  the  accompanying  table,  in  which  the  milligrams  of  fixed 
nitrogen  are  given. 

1.  Nutritive  medium  alone 2.6 

2.  Nutritive  medium  plus  watery  extract  of  ordinary  peat 1.9 

3.  Nutritive  medium  plus  watery  extract  of  fermented  peat 7.7 

4.  Alcoholic  extract  of  fermented  peat 6.5 

5.  Phosphotungstic  acid  precipitate  of  No.  4 5.6 

6.  Silver  fraction  of  No.  5 6.5 

We  may  see  from  these  results  that  the  above  bacteria,  having 
synthetic  ability  to  assimilate  nitrogen  from  the  air,  are  nevertheless 
influenced  by  vitamines.  While  the  vitamine  action  was  favorable 
to  the  fixation  of  nitrogen,  the  effect  upon  denitrifi cation  was  reduced, 
and  the  formation  of  ammonia  was  uninfluenced.  The  investigations 
were  then  applied  to  other  soil  bacteria  and  it  was  found,  as  a  rule, 
that  the  growth  of  the  nitrifying  bacteria  is  activated  by  vitamines, 
while  that  of  the  putrefaction,  denitrification,  and  ammonia  forming 
bacteria  is  not  affected.  Those  bacteria  which  have  the  power  of 
decomposing  organic  matter  should  in  fact  have  no  need  for  vitamines 
or  else  should  be  able  to  synthesize  them.  These  two  great  classes 
of  bacteria,  one  that  begins  the  nitrogen  cycle  and  the  other  that  ends 
it,  differ  from  each  other  entirely  in  their  metabolism  if  Mockeridge 
is  correct. 

It  must  be  especially  emphasized  that  Bottomley  and  his  school 
always  nioved  further  and  further  away  from  the  importance  of  the 
vitamines  for  plant  growth,  and  ascribed  this  remarkable  action  to 
the  nucleic  acid  derivatives.  These  conclusions  followed  in  the  paths 
which  Schreiner  and  Skinner  (187)  and  their  co-workers  laid  out 
for  the  action  of  nucleic  acid  derivatives  on  plant  growth.  These 
investigators  isolated  from  the  soil  some  purine  and  pyrimidine 
derivatives  and  tested  their  activity.  They  showed  that  xanthine 
could  increase  the  weight  of  wheat  sprouts  kept  in  water  about  21 
per  cent.  Similar  experiments  were  made  by  Macalister  (188)  with 
allantoine  and  by  Coppin  (189)  with  other  analogous  substances. 
Recently,  Miss  Mockeridge  (190),  experimenting  with  Lemna  major, 
showed  that  green  plants  need  an  addition  of  certain  other  sub- 
stances to  continue  to  live.  In  this  series,  the  influence  of  natural 


THE    VITAMINES   IN   THE   VEGETABLE   KINGDOM  71 

fertilizer  was  investigated.     The  results  obtained  in  the  fifth  week 
were  as  follows : 

Average  number 
of  plants 

1.  Control  solution 27.0 

2.  Extract  of  rotted  leaves 64.6 

3.  Extract  of  soil 64.4 

4.  Extract  of  fermented  peat 132. 6 

It  is  evident  that  identical  results  were  obtained  with  fertilizer 
and  with  bacterized  peat;  indeed,  fresh  fertilizer  was  less  active  than 
one  in  which  marked  bacterial  decomposition  had  set  in.  Mocke- 
ridge  then  found  that  nucleic  acids  and  nucleotides  predominated  in 
the  less  active  extracts,  while  in  the  more  active  extracts,  the  amount 
of  free  purine  and  pyrimidine  bases  was  increased.  Therefore,  the 
conclusion  was  made  that  the  effect  was  brought  about  not  by  the 
nucleic  acids  themselves,  but  by  their  decomposition  products. 
These  conclusions  are  obviously  not  justified  especially  since  the 
investigations  were  not  carried  out  with  pure  chemical  products; 
consequently  the  results  could  be  only  interpreted  to  indicate  that 
the  substrate  acted  favorably  on  plant  growth  because  of  some 
unknown  changes  that  had  taken  place  in  the  composition  of 
the  substrate.  Hydrolysis  of  the  afore-mentioned  nucleic  acids 
could  well  be  thought  of  as  a  secondary  occurrence  without  any 
physiological  significance.  It  is  very  likely  that  the  growth-promot- 
ing activity  of  peat  and  soil  extracts  is  not  due  to  the  nucleic  acid 
decomposition  products  but  to  the  presence  of  vitamines  which  are 
carried  down  in  the  fractionation.  Should  it  be  established  that  in 
the  growth  of  plants,  similar  substances  are  involved  as  in  the  growth 
of  yeast,  then  it  would  be  very  improbable  that  they  could  be  asso- 
ciated with  purine  and  pyrimidine  derivatives. 

Miss  Mockeridge's  idea  of  the  vitamine  cycle,  was  that  the  nitrify- 
ing bacteria  furnish  the  green  plants  with  the  growth-promoting 
substances  which  are  used  in  the  metabolism  of  the  plants,  partly 
transformed  into  the  vitamines  already  known  to  us,  and  which  can 
then  be  utilized  by  the  animals. 

The  investigations  of  Bottomley  and  Rosenheim  have  estab- 
lished that  higher  plants  also  need  a  vitamine  of  the  antiberiberi 
type  in  their  metabolism,  and  Bottomley  expressed  the  idea  that 
this  vitamine  occurred  in  the  plant  through  symbiosis  with  certain 
bacteria.  Whether  this  is  the  only  vitamine  that  plants  need, 


72  THE   VITAMINES 

remains  to  be  demonstrated.  Still,  it  is  apparent  that  the  vitamine 
of  type  A,  which  is  found  mostly  in  seeds  and  green  leaves,  is  formed 
in  the  leaves  perhaps  from  primary  products  which  must  be  furnished 
to  the  plants  from  some  outside  source.  The  question,  as  to  whether 
old  plants  that  already  have  leaves  need  extra  vitamine  in  the  same 
degree,  must  eventually  be  answered  and  may  perhaps  be  solved  by 
the  use  of  methods  for  sterile  growth,  as  proposed  by  Grafe  (191). 

Since  the  entire  animal  world  obtains  its  vitamine  requirements 
either  directly  or  indirectly  from  plant  food,  it  is  especially  important 
to  know  the  vitamine  cycle  in  its  entirety. 


CHAPTER   II 
THE  ROLE  OF  VITAMINES  IN  THE  ANIMAL  KINGDOM 

Coming  to  this  important  chapter,  we  must  first  of  all  satisfy  our- 
selves as  to  the- relative  importance  of  the  vitamines  in  nutrition. 
In  fact,  the  role  of  the  vitamines  is  placed  in  the  foreground  by  some 
investigators,  so  much  so,  that  the  impression  prevails  that  these 
substances  can  completely  compensate  for  all  dietary  deficiencies. 
Obviously,  this  is  not  the  case.  The  requirements  of  the  animal 
organism  for  each  of  the  food  constituents  have  been  determined 
chiefly  in  dogs,  and  lately  in  rats;  we  need  only  refer  the  reader  to 
the  modern  text  books  on  the  science  of  nutrition  if  he  wishes  to  study 
the  question  more  thoroughly.  We  must  specifically  emphasize,  as 
we  did  in  the  first  edition,  that  most  of  the  old  and  many  of  the  new 
experiments  on  this  subject  were  not  entirely  convincing,  since  the 
investigators  were  either  unaware  of  the  existence  of  the  vitamines  or 
else  ignored  their  significance.  In  the  last  six  years,  however,  many 
of  the  experiments  were  repeated,  bearing  in  mind  the  progress  that 
had  been  made  in  vitamine  research.  The  conclusions  arrived  at  in 
these  experiments,  for  example,  for  rats,  are  as  follows :  For  complete 
nutrition,  these  animals  need,  above  all,  a  biologically  complete 
protein — a  protein  of  high  biological  value — one  containing  all  of  the 
necessary  amino  acids;  further,  a  carbohydrate  of  the  nature  of 
starch,  sugar  or  dextrin;  a  certain  amount  of  fat,  a  salt  mixture,  so 
chosen  that  it  contains  the  necessary  cations  and  anions  in  proper 
proportion;  and  finally  vitamines  A  and  B.1  It  must  be  held  open 
for  further  investigation  to  determine  whether  or  not  rats  can  get 
along  on  a  complete  protein,  a  salt  mixture  and  vitamines  (without 
carbohydrate  and  fat) ;  for  some  animals,  like  the  dog  or  cat,  which 
can  get  along  on  meat  alone,  this  has  already  been  definitely  estab- 
lished. The  series  of  experiments  by  Spriggs  (192)  and  Maignon 

1  Parsons  (191a)  recently  found  that  rats  do  not  require  a  supply  of 
vitamine  from  outside  sources.  In  spite  of  the  lack  of  this  vitamine  in  the 
diet,  the  organs — spleen,  kidney  and  muscles — of  rats  so  fed  contain  appre- 
ciable amounts  of  vitamine  C  when  tested  on  guinea  pigs  in  the  form  of  extracts. 
Whether  this  vitamine  C  originates  from  the  vitamine  B  of  the  diet  has  to 
be  left  for  future  demonstration. 

73 


74  THE   VITAMINES 

(193),  dealing  with  the  harmful  effect  of  an  exclusive  protein  diet  on 
rats,  can  very  likely  be  attributed  to  the  insufficient  amount  of 
vitamines  supplied,  although  it  might  also  have  been  due  to  a  relative 
lack  of  salts.  In  a  special  case,  Maignon  (194)  noted  that  when  he 
fed  rats  on  egg  albumin,  together  with  some  salts,  they  developed 
a  disease  which  he  held  to  be  an  acute  intoxication  of  the  central 
nervous  system,  from  which  the  rats  died  in  a  few  days.  It  is  not 
improbable  that  it  was  some  sort  of  an  avitaminosis. 

Regarding  dietary  factors  such  as  nucleins,  lipoids,  etc.,  it  is  quite 
apparent  that  they  are  synthesized  in  the  animal  body.  It  is  often 
specified  that  for  certain  animals,  and  also  for  man,  the  animal 
proteins  are  preferable  to  the  vegetable  proteins.  In  practice  this 
is  quite  true,  and  it  may  be  explained  in  many  different  ways.  In 
the  first  place,  when  we  speak  ordinarily  of  animal  and  vegetable 
protein,  we  have  in  mind  the  natural  products  containing  those 
proteins.  Now,  there  are  great  fundamental  differences  between  the 
two  types  of  foods.  When  we  feed  an  animal  protein,  like  meat, 
eggs,  etc.,  we  give  the  protein,  which  is  very  poor  in  other  dietary 
components,  like  fat,  and  especially  carbohydrates,  in  very  concen- 
trated form.  With  plant  products  it  is  different;  they  contain 
protein  in  a  very  much  smaller  concentration,  accompanied,  for  the 
most  part,  by  large  amounts  of  carbohydrates.  That  this  may  be  of 
importance,  we  shall  see  later.  Furthermore,  it  is  possible  that  the 
amino  acids  of  animal  proteins,  because  of  their  close  resemblance 
in  composition  to  those  of  the  body  proteins,  are  better  utilized; 
besides  this,  they  may  contain  known  and  perhaps  even  unknown 
vitamines  carried  along  during  the  process  of  preparation.  For 
example,  Osborne,  Wakeman  and  Ferry  (I.e.  102)  found  that  it  is 
very  difficult  to  free  edestin  from  vitamine  B.  As  to  the  relative 
value  of  plant  and  animal  protein,  we  shall  go  into  it  more  fully  in 
discussing  human  nutrition. 

When  we  elaborated  the  vitamine  theory  in  1912,  we  stated  that 
these  new  substances  are  important  or  even  indispensable  to  all 
plants  and  animals,  predicated  upon  the  then  available  data.  The 
number  of  facts  which  we  can  bring  to  our  support  in  proof  of  the 
above  statement  has  been  greatly  augmented  since  the  first  edition 
of  this  book  appeared,  although  not  all  types  of  animals  have  been 
investigated  in  this  respect.  It  is  quite  true  that  some  animals 
placed  upon  a  vitamine-free  or  vitamine-poor  diet  do  not  remain 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM  75 

in  good  condition,  even  though  no  symptoms  of  any  of  the  known 
avitaminoses  are  apparent.  Nevertheless,  these  exceptions  cannot 
militate  against  the  general  significance  of  the  vitamines  for  the  life 
processes  for  many  reasons.  For  one,  so  long  as  the  experimental 
animals  are  kept  upon  an  artificial  diet  made  up  of  purified  con- 
stituents, they  do  not  live  long  if  extracts  of  unpurified  products 
are  not  given.  All  experiments  seeking  to  demonstrate  the  contrary 
have  been  carried  out  by  feeding  a  dietary  containing  either  traces 
of  known  or  unknown  vitamines.  Then  again,  it  has  also  been  shown 
that  some  species  of  animals  can  dispense  with  one  or  another  of  the 
vitamines.  This  phenomenon  may  perhaps  be  explained  by  the 
symbiosis  of  the  host  with  certain  bacteria.  In  the  higher  animals, 
these  microorganisms  are  found  in  the  intestinal  tract  in  large 
amounts  and  are  perhaps  able  to  build  up  the  vitamines  from  simple 
substances  and  convey  them  to  the  animal.  At  the  end  of  this 
chapter  we  shall  add  a  short  paragraph  dealing  with  the  role  of  bac- 
teria in  animals,  but  at  this  juncture,  we  shall  only  remark  that  no 
attempt  has  as  yet  been  made  to  establish  these  new  viewpoints 
experimentally. 

All  these  procedures  bring  us  nearer  to  the  point  that  interests  us 
mostly,  namely,  accurate  knowledge  of  the  subject  of  human  nutri- 
tion. At  once,  we  are  confronted  with  questions  dealing  with 
economic  and  agricultural  problems.  Let  us  consider  the  breeding 
of  animals  as  an  example.  From  everything  that  has  already 
been  mentioned  it  is  apparent  that  the  condition  of  the  soil,  which 
contains  quantities  of  organic  and  nitrogenous  matter,  can  be 
brought  into  relationship  with  the  study  of  vitamines.  The  indi- 
cation for  such  problems  is  seen  in  the  publication  of  W.  A.  Davis 
(195)  who  thought  that  the  poverty  of  the  soil  (in  Bihar,  India) 
in  phosphorus  is  the  cause  of  an  avitaminosis  in  cows,  which  is 
responsible  for  a  smaller  yield  of  milk  and  is  connected  with  the 
under-nourishment  of  children.  Similarly,  a  nervous  disease  of 
horses  was  observed  in  these  villages.  In  European  countries,  as 
well  as  in  the  United  States,  where  agriculture  and  cattle  raising  are 
carried  out  rationally,  these  conditions  are  of  course  to  be  noted 
very  infrequently.  It  is  different  in  countries  like  Australia,  Argen- 
tine and  South  Africa,  where  cattle  raising  is  carried  out  on  a  large 
scale  without  paying  sufficient  attention  to  the  care  of  the  soil,  due 
no  doubt  to  the  lack  of  laborers.  There  we  often  hear  of  diseases, 


76  THE   VITAMINES 

which  are  not  always  considered  as  deficiency  diseases,  though  they 
might  turn  out  to  be  such.  Because  of  insufficient  irrigation  of  the 
fields  and  periodic  dry  spells,  we  see  domestic  animals,  feeding  on 
wretched  vegetation  for  months  at  a  time,  dying  from  diseases  which 
we  shall  describe  more  in  detail  further  along  in  the  chapter.  Here, 
we  are  perhaps  dealing  with  conditions  which  follow  each  other 
like  the  individual  links  of  a  chain.  As  a  result  of  the  unfavorable 
condition  of  the  soil  and  the  paucity  of  soil  bacteria,  the  vegetation 
suffers.  The  animals  which  must  feed  on  this  vegetation  either 
become  sick,  or  else  are  not  in  perfect  health,  though  no  unusual 
symptoms  may  be  apparent.  Subsequently,  their  offspring  and 
the  children  drinking  the  milk  of  such  animals  are  naturally  poorly 
nourished;  the  same  is  true  of  grown  people  who  use  the  meat  of 
such  animals.  Indeed,  these  conditions  could  as  well  prevail  in 
European  countries  where,  because  of  the  war,  the  number  of  cattle 
has  decreased,  leading  to  the  exhaustion  of  natural  fertilizer,  which 
could  not  be  replaced  by  artificial  fertilizer  because  of  the  difficulty 
of  obtaining  it.  The  diagram,  illustrative  of  the  relationship  be- 
tween the  above  mentioned  factors,  is  shown  below. 

/Unfavorable  soil  conditions 
\ 

Lack  of  soil  bacteria  Insufficient  fertilizer 

I  T 

Wretched  vegetation >      Diseased  animals 

I 

Meat  and  milk  of  incomplete 
nutritive  value. 
Avitaminoses  in  man 

LIFE   WITHOUT   BACTERIA 

As  we  have  stated  several  times  previously,  the  incomplete  knowl- 
edge of  the  role  of  the  bacterial  flora  in  animals 'is  the  cause  of  a 
great  gap  in  the  conception  of  the  general  significance  of  the 
vitamines.  We  know,  or  at  least  we  suppose,  that  some  bacteria 
are  independent  in  their  metabolism  of  an  outside  addition  of  vita- 
mine,  and  since  we  know  that  a  great  number  of  bacteria  live  in  the 
intestinal  canal,  either  in  symbiosis  or  else  as  a  parasite,  we  can 
easily  imagine  that  their  functions  may  have  something  to  do  with 
supplying  the  host  with  vitamines. 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  77 

Long  before  the  introduction  of  the  vitamine  theory  in  the  study 
of  nutrition,  Schottelius  (196)  sought  to  solve  the  problem  as  to 
whether  it  was  possible  to  raise  animals  in  a  sterile  state,  and  to  keep 
them  alive  for  a  long  time.  After  overcoming  great  experimental 
difficulties,  among  which  was  the  construction-  of  specially  designed 
apparatus,  he  wa's  finally  able  to  raise  and  keep  young  chickens  under 
sterile  conditions.  The  animals,  however,  soon  became  cachectic  and 
died.  Using  similar  methods,  Belonowsky  (197)  was  no  more  fortu- 
nate. The  problem  was  then  undertaken  with  lower  and  higher 
animals  by  various  workers.  Of  the  lower  animals,  flies  were  chiefly 
used.  In  this  connection,  there  are  the  experiments  of  Bogdanow 
(198)  who  attempted  to  raise  larvae  of  meat  flies  (Calliphora  vomi- 
toria)  in  a  sterile  condition.  He  proceeded  as  follows :  the  eggs  were 
sterilized  with  sublimate  and  the  larvae  cultivated  upon  sterile  meat. 
They  grew  very  slowly,  and  showed  no  metamorphosis.  Since  they 
did  not  survive,  Bogdanow  concluded  that  bacteria  are  necessary  for 
this  type  of  insect.  In  a  number  of  experiments,  Delcourt  and 
Guyenot  (199),  and  also  Guye*not  himself,  sought  to  reach  the  goal 
by  the  use  of  a  different  method.  They  attempted  to  obtain  aseptic 
larvae  through  successive  cultures  on  an  acid  medium.  They  noted 
that  flies  could  live  on  yeast  and,  after  several  generations,  also  on 
sterile  potato.  Larvae  grew  well  on  peptone  and  salt,  but  showed 
no  metamorphosis.  Wollmann  (200),  following  the  method  of 
Bogdanow,  carried  these  investigations  further,  and  came  to  the 
same  conclusions  as  the  latter. 

A  number  of  animals,  like  the  scorpion  and  winter  lizard,  are 
supposed  to  have  sterile  intestines  and  hence  are  specially  suitable 
for  such  experiments.  Mme.  Metchnikoff  (201)  tried  to  raise  frogs' 
larvae  in  a  sterile  state  but  without  success.  Moro  (202)  had  already 
made  similar  experiments  with  larvae  of  the  alliaceous  toad.  The 
larvae  could  be  kept  alive  for  35  days,  but  their  growth  was  much 
less  favorable  than  that  of  the  controls.  Since  all  these  experiments 
appeared  to  be  rather  indefinite  (at  least  for  the  pre-vitamine  period), 
Cohendy  (203)  undertook  the  experiments  of  Schottelius  with 
chickens.  For  this  purpose,  he  built  a  very  ingenious  apparatus,  in 
which  the  food,  together  with  the  whole  paraphernalia,  was  sterilized 
at  115°C.  for  25  minutes,  or  for  1J  hours  at  118°C.  Cohendy 
reported  that  he  had  obtained  good  results  with  this  arrangement, 
and  came  to  the  conclusion  that  life  is  quite  possible  in  the  absence 


78  THE   VITAMINES 

of  bacteria.  Upon  reading  through  his  work,  one  receives  the 
impression  that  although  the  animals  ate  more  food  than  the  controls, 
they  weighed  much  less,  and  soon  died.  The  longest  duration  of  life 
was  40  days  (204).  Schottelius  (205),  criticising  the  above  work, 
expressed  the  idea  that  Cohendy's  experiments  showed,  on  the  con- 
trary, that  the  intestinal  bacteria  were  useful,  or  even  indispensable. 
Such  experiments  were  likewise  carried  out  with  mammals. 
Ktister  (206),  working  with  goats,  was  able  to  maintain  young 
animals  in  a  sterile  condition  for  35  days,  during  which  time  no 
difference  from  the  controls  was  noted.  It  is  not  impossible  that 
these  experiments  did  not  last  long  enough.  Charrin  and  Guille- 
monat  (207)  carried  out  similar  experiments  on  guinea  pigs,  while 
Kianizin  (208)  used  guinea  pigs  and  rabbits.  The  latter  made  his 
animals  breathe  sterilized  air  and  eat  sterilized  food.  The  poor 
results  of  this  regime  were  noticeable  in  from  4  to  9  days;  the  animals 
suffered  from  weakness  and  died  soon  thereafter.  Nutall  and  Thier- 
felder  (209)  conducted  the  well  known  experiments  on  guinea  pigs 
in  which  these  animals  were  kept  free  from  bacteria  and  symptoms 
of  sickness  and  even  showed  a  gain  in  weight.  In  this  case,  too,the 
experiments  were  of  rather  short  duration. 

These  experiments  required  corroboration  from  the  viewpoint  of 
progress  in  vitamine  research.  This  view  is  held  likewise  by  Weill 
and  Mouriquand  (210)  and  Schaeffer  (211).  It  must  be  pointed 
out  that  while  all  the  investigators  working  on  the  problem  of  bac- 
teria-free existence  thought  that  they  were  dealing  with  only  a 
single  unknown  factor  (the  role  of  bacteria),  there  were  really  two 
to  be  considered.  During  the  process  of  food  sterilization,  the 
vitamines — according  to  the  choice  of  food  and  the  conditions  of 
temperature — were  more  or  less  affected  by  the  heat,  and  probably 
destroyed.  It  is  obvious,  therefore,  that  the  problem  of  a  germ- 
free  existence  was  much  more  complicated  than  was  imagined  by  the 
above  mentioned  investigators.  From  the  facts  known  at  present, 
we  may  conclude  that  in  some  animals,  in  which  it  is  not  possible  to 
produce  an  avitaminosis  by  means  of  a  diet  either  poor  or  lacking  in 
vitamines,  the  bacterial  intestinal  flora  possibly  assume  the  task  of 
preparing  the  necessary  substances  for  the  host.  To  solve  the  ques- 
tion beyond  all  doubt,  it  is  necessary  to  proceed  so,  that  in  the 
first  place  we  arrange  for  a  bacteria-free  growth;  if  this  should  fail, 
the  animals  may  perhaps  be  saved  through  the  addition  of  vita- 


THE    VITAMINES   IN   THE    ANIMAL   KINGDOM  79 

mine.  It  may  then  be  possible  to  experiment  with  the  pure  cul- 
tures of  the  intestinal  bacteria  indigenous  to  the  particular  species, 
to  produce  a  normal  dietetic  condition  without  the  addition  of  vita- 
mine.  Only  then  shall  we  be  in  a  position  to  state  that  vitamines 
are  necessary  for  the  existence  of  the  entire  animal  kingdom  and 
eventually  for  the  whole  organized  world.2  Sometimes,  however, 
a  poor  idea  has  some  truth  in  it,  and  this  seems  to  be  the  case  with 
Portier 's  hypothesis.  It  is  possible,  as  we  have  already  said,  that 
certain  animals  and  plants  obtain  their  vitamines  through  a  sym- 
biosis with  bacteria.  This  idea  was  tested  experimentally  by 
Portier  and  Random  (217).  They  made  the  observation  that 
rabbits  fed  on  sterilized  cabbage  and  carrots  developed  an  avitamin- 
osis,  while  rabbits  which  received  an  addition  of  the  excrement  of 
another  rabbit  on  the  same  diet  remained  normal;  still,  it  is  not 
quite  clear  to  us  why  rabbits  kept  on  the  same  diet  should  possess 
a  different  intestinal  flora  than  those  receiving  an  addition  of  feces. 
It  appears  that  these  experiments,  serving  to  demonstrate  the 
importance  of  the  symbiotic  intestinal  bacteria,  could  be  much  better 
explained  on  the  basis  of  the  vitamine  content  of  the  feces. 

PROTOZOA 

Investigations  on  protozoa  were  carried  out  chiefly  with  paramecia. 
Lund  (218)  has  recently  shown  that  he  grew  these  single-cell  organ- 
isms on  yeast  extract,  but  the  details  of  the  experiment  have  not  yet 
been  published.  Calkins  and  Eddy  (219)  worked  with  Paramecium 
aurelia  kept  on  two  different  nutritive  media — hay  infusion  and 
cooked  flour.  As  a  source  of  vitamine,  a  preparation  of  vitamine  B 
of  the  pancreas  was  used.  The  number  of  daughter  cells  varied 
within  the  limits  of  experimental  error,  and  therefore  it  was  con- 
cluded that  the  vitamine  exerted  no  influence  on  this  type  of  animal. 
Flather  (220)  obtained  somewhat  dissimilar  results  from  experiments 
in  which  pure  cultures  of  species  of  paramecia  were  maintained 
always  under  the  same  experimental  conditions;  in  every  experiment, 

2  Portier  (212)  has  recently  formulated  a  hypothesis  in  which  he  assumes 
that  in  certain  plant  and  animal  organs,  there  are  heat  resisting  bacteria 
(called  symbiotes)  which  behave  biochemically  like  vitamines  (Bierry  and 
Portier  (213).  Just  now,  we  shall  not  discuss  this  theory  itself  for  it  appears 
to  be  lacking  a  real  basis,  and  besides  it  has  already  been  made  the  subject 
of  an  earnest  critique  by  Delage  (214),  Lumiere  (215)  and  Ranc  (216). 


80  THE    VITAMINES 

three  cells  were  grown  in  two  drops  of  water,  with  the  addition  of  one 
drop  of  a  1  per  cent  watery  solution  of  polished  or  unpolished  rice, 
or  malted  milk.  The  division  factor  was  taken  as  a  measure  of  cell 
activity  and  the  first  24  hours  were  disregarded,  to  give  the  cells 
the  opportunity  of  adjusting  themselves  to  the  new  conditions.  The 
following  values  were  obtained:  for  white  rice  0.34,  for  unpolished 
rice  0.58,  for  malted  milk  0.84.  It  was  then  possible,  by  the  addition 
of  malted  milk,  to  make  the  nutritive  value  of  the  white  rice  approxi- 
mate that  of  the  malted  milk  Other  experiments  were  made  with 
various  concentrations  of  malted  milk  and  also  with  orange  juice, 
with  the  result  that  an  addition  of  malted  milk  in  higher  concentra- 
tions was  of  no  further  advantage,  while  orange  juice  showed  a  very 
slight  activity  Addition  of  orange  juice  to  white  rice  was  followed 
by  better  results  than  when  added  to  unpolished  rice,  and  with 
orange  juice  alone,  no  paramecia  cells  could  live  after  six  days.  In 
these  experiments,  the  influence  of  vitamines  is  easily  recognized; 
in  fact,  Flather  was  undoubtedly  dealing  in  this  case,  not  with  one 
vitamine  but  with  the  combined  influence  of  the  mixture  of  vitamines. 

Chambers  (221)  carried  out  similar  experiments  which  demon- 
strated the  slight  effect  of  potato  extract ;  this  displayed  some  influ- 
ence only  if  the  nutritive  solution  did  not  contain  sufficient  dietary 
constituents;  the  same  was  true  of  malted  milk.  Peters  (222) 
cultivated  Colpidium  colpoda  on  a  nutritive  solution  which  con- 
tained, besides  salts,  a  mixture  of  some  amino  acids  and  in  some 
cases  only  one  amino  acid,  tryptophane.  The  mixture  of  amino 
acids  was  more  active  than  the  tryptophane  alone.  The  first 
divisions  took  place  very  slowly,  though  it  was  possible  to  keep  the 
cultures  alive  for  three  months  through  sub-culture.  Peters  ex- 
plicitly emphasized  that  he  could  find  no  symbiotic  microorganism. 
He  stated  that  in  certain  stages  of  development,  a  vitamine-like 
substance  seems  to  play  a  definite  role.  In  all  these  experiments, 
having  for  the  purpose  the  study  of  the  influence  of  the  vitamines, 
it  is  important  that  the  food  mixture  used  should  be  proved  to  be 
vitamine-free.  Abderhalden  and  Kohler  (I.e.  127)  investigated 
the  action  of  yeast  extracts  on  flagellates  (colpoda)  with  positive 
findings. 

When  we  leave  the  protozoa,  we  find  a  dearth  of  available  work 
on  other  kinds  of  animals,  although  it  should  not  be  thought  that 
such  work  does  not  exist.  This  is  specially  true  of  the  polycellular 


THE    VITAMINES   IN   THE    ANIMAL    KINGDOM  81 

animals  investigated  at  various  biological  experimental  stations, 
where  observations  which  would  be  of  interest  to  us  have  undoubtedly 
been  made  on  the  modes  of  nutrition  of  many  insects.  Unfortunately, 
these  data  were  unavailable  to  us. 

METAZOA 

Coming  to  the  polycellular  animals,  we  shall  first  describe  a  series 
of  experiments  undertaken  for  some  other  purposes.  They  deal  with 
the  influence  of  lecithin  upon  growth  and  were  carried  out  by  Goldfarb 
(223).  In  this  respect,  he  investigated  a  whole  series  of  animals,  and 
also  tadpoles  and  eggs  of  the  starfish.  At  first,  he  stated  emphati- 
cally that  lecithin  had  a  definite  action,  but  in  a  later  work  (224), 
he  admitted  that  it  was  inactive.  This  is  apparently  associated 
with  a  vitamine  impurity,  whose  presence  may  vary  with  the  mode 
of  preparation  of  lecithin. 

It  may  not  be  amiss  to  say  a  few  words  here  on  Carrel's  tissue 
culture  in  vitro,  which  in  t-he  future  may  perhaps  be  of  help  in  advanc- 
ing the  vitamine  problem.  These  tissue  cultures  may  be  thought  of 
as  polycellular  organisms. 

The  growth  of  tissue  in  vitro 

Most  of  the  experiments  of  Carrel  were  made  at  a  time  when  the 
vitamines  were  either  unknown  or  given  very  little  consideration; 
otherwise  it  might  have  been  possible  to  choose  still  better  conditions 
for  tissue  culture.  We  must  admit,  however,  that  Carrel  practically 
foresaw  the  future  trend  of  work,  in  that  he  chose  the  natural  juices 
of  the  animal  organism  as  the  nutritive  solutions.  Inasmuch  as  the 
author  has  little  practical  knowledge  of  this  method,  he  is  naturally 
not  in  a  position  to  judge  whether  or  not  Carrel  would  have  obtained 
even  better  results  if  he  had  tested  his  nutritive  solutions  for  the 
presence  of  all  the  necessary  elements  for  nutrition.  Because  of  the 
great  interest  that  experimental  surgery  of  the  future  may  have  in 
this  method,  it  is  very  important  to  obtain  the  greatest  success. 
The  tissue  fragments  used  in  these  experiments,  are  removed  from 
their  normal  environment  and  must  then  lead  an  independent  exist- 
ence. For  this  purpose,  they  need  all  of  the  dietary  constituents 
(also  vitamines)  designated  as  essential  by  the  modern  science  of 


82  THE   VITAMINES 

nutrition.  By  means  of  frequent  sub-cultures,  Carrel  prevented  the 
nutritive  substances  from  becoming  exhausted.  That  this  had  to  be 
resorted  to  frequently  in  order  to  keep  a  fragment  of  tissue  alive  for 
a  long  while,  shows  perhaps  that  the  conditions  for  growth  could 
have  been  better  chosen. 

In  this  respect,  the  method  may  be  applicable  to  a  new  field  of 
work.  We  have  in  mind  particularly  Carrel's  antiseptic  method 
of  treating  wounds,  which  came  into  use  during  the  war.  It  is 
apparent  that  the  organism  suffering  from  shock  and  other  complica- 
tions could  not  be  in  the  best  nutritive  condition;  and  in  the  wounded 
tissue  itself,  through  the  disturbance  of  the  blood  circulation,  con- 
ditions could  not  be  the  most  favorable.  For  these  reasons,  it  would 
be  of  interest  to  apply  to  the  treatment  of  the  wound,  those  facts 
that  have  been  learned  in  the  study  of  the  composition  of  nutritive 
fluids  best  suited  for  the  tissues.  Practically,  this  could  be  brought 
about  by  the  substitution  of  the  antiseptic  solution  by  a  nutritive 
solution  from  time  to  time,  with  the  possibility  that  the  wounded 
tissue  could  be  nourished  and  the  process  of  healing  hastened. 

The  method  of  Carrel  (225)  is  as  follows :  Under  appropriate  con- 
ditions, tissue  fragments,  and  pieces  of  tumors  too,  may  be  grown  for 
more  than  four  months,  during  which  the  tissue  shows  growth  and 
peripheral  expansion.  The  tissue  (226),  antiseptically  prepared,  is 
placed  in  a  plasma  diluted  with  J  to  f  of  its  volume  of  distilled 
water.  For  this  purpose,  it  is  best  to  use  the  plasma  of  the  animal 
under  investigation  or  that  of  a  homologous  animal.  In  from  3  to 
4  days,  the  tissue  culture  (Carrel  (227),  Carrel  and  Burrows  (228)) 
is  placed  for  a  few  minutes  in  Ringer's  solution  and  then  transferred 
to  a  fresh  plasma.  Some  tissue  cultures  have  been  kept  alive  for 
four  and  a  half  months,  having  been  transferred  as  above  48  times. 
A  fragment  of  heart  continued  to  beat  after  140  days.  Only  embry- 
onic tissues  and  tumors  could  grow  in  an  artificial  medium  (Locke's 
solution,  agar  and  bouillon).  In  a  later  work,  Carrel  (229)  investi- 
gated in  greater  detail  the  influence  of  tissue  extracts  and  body  fluids 
(as  did  Walton  (230)  too),  and  found  that  they  increased  the  growth 
of  connective  tissue  in  particular  from  3  to  40  times.  This  was  the 
case  especially  with  embryonic  spleen  extracts,  and  it  was  consider- 
ably smaller  with  greater  dilutions.  It  was  true  only  of  the  same 
type  of  animal,  and  the  action  of  the  extracts  was  weaker  on  heating 
to  56°C.  and  entirely  dissipated  at  70°C.  The  active  substance 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM  83 

could  be  filtered  through  a  Berkefeld  filter,  but  not  through  a  Cham- 
berland  filter.  Regarding  the  foregoing,  it  appears  likely  that  a  new 
field  for  vitamine  research  is  opened  up. 

INSECTS 

We  have  to  deal  here  with  the  very  interesting  studies  of  Jacques 
Loeb  (231).  He  demonstrated  that  a  type  of  fly  (Drosophila)  could 
attain  the  larva  stage  upon  a  solution  of  cane  sugar  and  salts,  together 
with  the  addition  of  filter  paper.  The  larva?  grew  quickly,  especially 
on  the  addition  of  alanine  or  an  ammonium  salt.  In  this  particular 
instance,  no  attention  was  paid  to  the  influence  of  the  bacteria  on  the 
substrate.  Five  successive  generations  of  the  banana  fly  (232)  were 
grown  on  a  solution  of  glucose,  cane  sugar,  ammonium  tartrate,  citric 
acid,  di-potassium  phosphate  and  magnesium  sulphate.  As  Loeb 
himself  observed,  it  was  not  quite  certain  that  the  solution  was  free 
of  bacteria;  the  presence  of  yeast  in  particular  was  not  excluded. 
In  these  experiments,  the  flies  themselves  were  not  sterile  and  there- 
fore Loeb  and  Northrop  (233)  started  experiments  with  flies  hatched 
from  eggs  sterilized  by  a  sublimate  solution.  Twelve  sterile  genera- 
tions were  cultivated  in  this  manner,  carefully  protected  from  every 
possibility  of  contamination  with  yeast  growth.  The  nutritive  solu- 
tion used  was  composed  of  450  grams  baker's  yeast  and  50  grams 
citric  acid  (used  for  the  purpose  of  decreasing  the  danger  of  infection 
during  the  various  manipulations)  in  a  liter  of  water.  In  a  great 
number  of  experiments,  the  investigators  showed  that  on  another 
medium,  for  instance,  filter  paper  and  cane  sugar,  with  the  addition 
of  salts,  and  likewise  with  the  addition  of  casein,  edestin,  egg  albumin, 
milk,  or  a  mixture  of  amino  acids,  the  larvae  attained  normal  size 
but  could  not  be  brought  to  the  stage  of  metamorphosis.  Sterile 
flies,  grown  on  sterile  bananas  or  potatoes,  showed  no  sexual  develop- 
ment. The  results  with  yeast  as  substrate  were  different;  here,  all 
the  substances  are  manifestly  at  hand,  which  are  indispensable  for 
the  growth  and  development  of  flies.  Butter,  nucleic  acids,  thymus 
and  thyroid  extracts  were  without  influence,  and  the  authors  believed 
that  these  substances  are  different  from  those  which  are  of  importance 
for  pigeons,  rats  and  other  warm-blooded  animals.  This  view  must 
naturally  undergo  a  slight  modification,  since  there  is  obviously  no 
reason  to  differentiate  between  flies  and  warm-blooded  species.  The 


84  THE    VITAMINES 

yeast  substance  was  heat-resistant  but  lost  some  of  its  activity  on 
treatment  with  cold  or  hot  alcohol.  Drosophila  is  a  monophageous 
animal  and  yeast  seems  to  be  the  only  adequate  food  for  it.  These 
flies  can  live  in  any  nutritive  solution  that'  is  suitable  for  the  growth 
of  yeast,  and  the  medium  is  usually  infected  with  this  organism. 
Loeb  and  Northrop  rightly  observed  that  in  considering  the  synthetic 
abilities  of  the  higher  animals,  one  must  not  lose  sight  of  a  possible 
cooperation  of  the  intestinal  bacteria.  Loeb  and  Northrop  (234)  and 
also  Northrop  (235)  showed  by  means  of  a  specially  planned  series 
of  experiments  that  the  developmental  stages  of  flies  (except  imago) 
can  be  prolonged  from  8  to  17  days  at  will.  They  believed  that  the 
three  stages  of  metamorphosis  could  be  regulated  by  the  formation 
and  disappearance  of  three  different  substances. 

In  a  later  work,  Northrop  (236)  investigated  the  role  of  yeast  in 
the  nutrition  of  flies.  He  found  that  the  number  of  flies  that  could 
be  developed  on  a  certain  quantity  of  yeast  could  be  increased  by 
the  addition  of  bananas,  casein  or  sugar.  The  rate  of  growth  on  a 
combination  of  yeast  and  bananas  is  greater  than  on  yeast  alone. 
In  mixtures  containing  a  small  amount  of  yeast,  growth  proceeds 
more  slowly,  and  in  a  dilution  of  1  to  128,  it  becomes  entirely  abnormal. 
Kidney,  liver,  dog  pancreas,  mouse  liver,  and  bodies  of  flies  them- 
selves were  suitable  for  the  growth  of  the  larvae,  while  sterilized 
spleen,  heart  muscle,  suprarenal  glands,  thyroid  and  blood  of  the 
dog  were  unsuitable;  the  addition  of  muscle  tissue,  testicles  and 
sterilized  thymus  gland  of  the  dog,  rabbit  and  calf,  resulted  in  the 
development  of  some  chrysalis,  but  growth  proceeded  slowly  and 
the  flies  appeared  dwarfed.  Robertson's  tethelin  (lipoid  from  the 
hypophysis)  was  inactive. 

Guyenot  (237)  attributed  to  the  vitamines  some  part  in  the  devel- 
opment of  fly  larvae  of  the  Drosophila  ampelophila.  Baumberger 
(238)  investigated  some  species  of  flies  and  came  to  the  conclusion 
that  Drosphila  melanogaster  can  live  on  fermenting  fruits  and  also  on 
yeast  protein.  A  combination  of  yeast,  nucleo-proteids,  sugar  and 
a  salt  mixture  provides  a  suitable  food  for  larvae.  Other  kinds  of 
flies,  like  Musca  domestica,  Desmotopa,  Sciaria  and  Tyroglypha,  can 
all  thrive  on  microorganisms. 

From  this  chapter,  we  see,  especially  from  the  work  of  Loeb,  that 
flies  need  vitamines  for  their  development,  and  that  vitamine  B 
plays  the  most  important  part. 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM  85 

FISH 

As  we  progress  in  the  animal  series,  we  do  not  meet  with  any 
vitamine  experiments — at  least  to  our  knowledge — till  we  come  to 
fish,  the  single  great  group  typical  of  the  vertebrates,  of  which  we 
have  no  knowledge  in  this  respect.  There  is  only  the  work  of  Mor- 
gulis  (239)  which  might  be  considered  to  have  some  slight  relationship 
to  our  subject.  He  fed  ox  heart  to  fish  and  found  that  it  is  better 
assimilated  when  cooked,  while  ox  liver  had  no  particular  peculiari- 
ties. Almy  and  Robinson  (240),  comparing  dry  and  fresh  fish  food, 
found  that  with  the  latter  there  was  less  mortality  and  a  more  definite 
weight  increase. 

AMPHIBIA 

Emmett  and  Allen  (241)  used  frog  larvae  in  their  vitamine  studies, 
but  the  fact  that  the  larvae  consumed  each  other  complicated  the 
experimental  work  somewhat.  The  larvae  reacted  poorly  towards  a 
greater  addition  of  fat,  and  developed  better  on  a  diet  containing  5 
per  cent  fat  than  on  one  containing  28  per  cent.  Both  vitamines,  A 
and  B,  appeared  to  be  necessary,  although  the  latter  seemed  more 
important.  The  variation  of  protein  content  from  10  to  30  per  cent 
was  of  no  special  significance,  although  the  optimum  growth  was 
obtained  with  higher  concentrations.  Lactalbumin  or  a  mixture  of 
beef  and  oats  was  superior  to  corn  gluten,  indicating  the  importance 
of  the  protein  used.  The  growth  of  the  hind  legs  depends  upon 
the  addition  of  vitamine,  according  to  the  findings  of  Emmett,  Allen 
and  Sturtevant  (242) .  For  complete  metamorphosis,  the  addition  of 
iodine  also  appears  to  be  necessary. 

Harden  and  Zilva  (243)  investigated  both  larvae  and  adult  frogs. 
With  tadpoles  kept  in  water  to  which  was  added  fresh  meat,  a  normal 
development  was  noted,  while  an  experimental  diet  made  up  of  20 
per  cent  casein,  75  per  cent  starch,  5  per  cent  salts  with  the  addition 
of  butter,  yeast  extract,  and  orange  juice,  did  not  permit  of  normal 
development,  since  only  two  out  of  six  animals  attained  full  growth. 
According  to  this  a  better  experimental  diet  had  to  be  prepared. 
With  adult  frogs,  another  difficulty  presented  itself  in  the  develop- 
ment of  a  fungus-like  disease.  On  a  diet  free  from  vitamine  B,  some 
of  the  animals  stretched  out  their  legs,  before  death,  and  one  suffered 
from  convulsions.  Without  A-  and  B-vitamines,  seven  frogs  died  in 
about  four  months.  Out  of  five  animals  on  a  diet  containing  the 


86  THE    VITAMINES 


PCR/0D 


f1  ft  ff  ft  ft 

Hi     12  /  I  I 


FIG.  6.  GROWTH  OF  FROG  LARVAE 

Diet  Hi  contained  A-  and  B-vitamines,  while  diet  112  was  lacking  in  B- 
vitamine   (Emmett-Allen). 


FIG.     5 


FIG.  7.  GROWTH  OF  FROG  LARVAE 

Diet    Ilia    was    complete    while    diet    VI2    was    deficient    in    vitamines 
(Emmett-Allen). 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  87 

above  two  vitamines,  with  or  without  vitamine  C,  only  one  died. 
Out  of  six  animals  getting  A-vitamine  but  not  C-vitamine,  all  died 
in  about  155  days.  Out  of  five  animals  receiving  vitamine  B,  with 
or  without  C,  four  survived  for  250  days. 

Accordingly,  B-vitamine  is  necessary  for  the  maintenance  of 
grown  frogs,  although  the  symptoms  were  noticeable  only  after  six 
months,  while  the  C-vitamine  appears  to  exert  no  influence  on  the 
animals. 

BIRDS 

Chickens 

As  stated  in  the  historical  part,  Eijkman  (I.e.  50)  and  Grijns  (I.e. 
52)  were  the  first  to  experiment  with  chickens.  It  was  particularly 
noted  that  Eijkman,  by  feeding  chickens  with  the  food  discarded  by 
a  beriberi  hospital,  observed  that  they  developed  a  disease  which  he 
called  "Polyneuritis  gallinarum."  Eijkman  soon  found  that  other 
foodstuffs,  like  sago  and  starch,  could  also  bring  on  this  disease. 

Normal  nutrition  of  chickens.  We  shall  first  consider  the  normal 
dietary  requirements  of  the  chicken.  One  who  has  tried  to  raise 
young  chickens  in  the  laboratory  knows  the  great  difficulties  that 
have  to  be  dealt  with.  The  first  attempt  in  this  direction  was  made 
by  the  author  (244)  who  investigated  the  influence  of  various  kinds  of 
foods  on  the  growth  of  experimental  tumors.  In  spite  of  the  great 
eagerness  with  which  the  chickens  ate  the  food,  growth  ceased  and 
they  developed  a  characteristic  disease  of  the  legs;  the  toes  lost  their 
vigor  and  were  bent.  Very  often  it  was  noticeable  that  the  beak  was 
not  closed  properly,  and  the  animals  died  because  of  their  inability 
to  pick  up  food.  The  eyes  were  frequently  closed,  and  the  chickens 
suffered  from  a  disease  which  gradually  led  to  complete  blindness, 
and  which  would  have  been  regarded  to-day  as  ophthalmia.  A 
change  in  the  diet  was  not  enough  to  better  conditions  since  the 
animals  were  already  receiving  quite  a  variety  of  food.  An  attempt 
was  made  to  feed  the  chickens  with  live  grub-worms,  and  for  a  time 
the  floors  of  the  cages  were  covered  with  grass  and  earth,  which  was 
frequently  renewed.  We  believed  then  that  the  wooden  floors  had 
some  pathological  effect  on  the  legs  of  the  animals.  However,  all 
these  measures  were  of  no  avail  and  the  birds  became  sick,  showing 
symptoms  which  we  then  attributed  to  rickets.  On  this  basis,  we 
used  cod  liver  oil  as  a  therapeutic  measure.  As  the  figures  below 
indicate,  a  young  chicken  could  maintain  itself  for  eight  months  on 


88 


THE    VIT AMINES 


unpolished  rice  and  cod  liver  oil  (Funk  and  Macallum  245).  The 
animal  was  normally  built  but  weighed  only  160  grams  while  a  chicken 
of  this  age  should  weigh  about  two  kilos.  No  symptoms  that  could 
have  been  regarded  as  rachitic  were  noted. 

The  animal  had  a  large  beak  and  excessively  long  feathers  for  his 
size;  all  secondary  sexual  indications  were  lacking  and  it  chirped  like 
a  four  weeks  old  chick. 

On  white  rice,  the  young  chickens  died  (I.e.  244)  after  about  two 
weeks,  while  on  unpolished  rice  it  was  possible  to  keep  them  alive  for 
more  than  5  weeks.  When  the  young  chickens  were  fed  Spratt's 
chicken  food,  the  weight  increased  more  than  twice,  while  on  unpol- 
ished rice  for  4  weeks,  there  was  no  growth  at  all.  These  experi- 


FIG.  8.  SEVEN  MONTHS  OLD  CHICKENS 

Normal  diet  and  a  diet  of  unpolished  rice  and  cod^  liver  oil.  (Funk -Mac- 
allum). Left— 2500  grams;  right— 160  grams. 

ments  were  conducted  with  15  days  old  chickens  and  were  later 
repeated,  with  the  same  results  (Funk  246).  In  all  exact  feeding 
experiments  with  chickens,  it  is  necessary,  particularly  during  the 
summer,  to  cover  the  cages  with  muslin,  otherwise  they  eat  so  many 
flies  that  the  results  may  be  misleading.  We  have  often  observed 
that  a  chicken  which  grew  in  spite  of  a  deficient  diet  stopped  growing 
promptly  as  soon  as  the  cage  was  covered. 

Drummond  (247) 3  carried  our  experiments  further  and  criticized 
our  results  rather  sharply.  He  found  that  the  possibility  of  raising 

3  After  my  departure  from  the  London  Cancer  Hospital  Research  Institute 
in  1915,  my  former  assistant,  J.  C.  Drummond,  continued  a  great  number  of 
experiments  started  by  me,  without  communicating  with  me,  and  without 
stating  in  his  publications  that  the  ideas  had  originated  with  me. 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  89 

chickens  in  the  laboratory  to  full  growth  depended  upon  the  age  of 
the  chickens  used.  He  observed  that  the  animals  often  failed  to  grow 
on  a  normal  diet,  and  he  accordingly  believed  that  this  suppression 
of  growth  had  nothing  to  do  with  the  vitamines,  although  his  results 
with  white  and  with  red  (unpolished)  rice,  as  compared  with  a  normal 
diet,  were  identical  with  ours.  Drummond  then  tried  to  improve 
the  condition  of  the  animals  by  giving  them  earth,  sand  and  wood 
chips,  but  without  success.  We  showed  later  (248)  that  it  was 
possible  to  decrease  the  mortality  of  the  animals  and  to  improve 
their  general  condition,  by  adding  animal  charcoal  to  the  diet  and 
substituting  milk  for  the  drinking  water. 

Buckner,  Nollau  and  Kastle  (249)  demonstrated  that  young 
chickens  did  not  grow  well  on  a  diet  poor  in  lysine,  as  compared  to 
one  rich  in  lysine;  the  addition  of  butter  was  without  effect.  It  is 
not  improbable  that  the  diet  rich  in  lysine  possessed  altogether  better 
nutritive  value.  Similar  results  with  lysine  were  obtained  by 
Osborne  and  Mendel  (250).  In  a  later  investigation,  they  (251) 
occupied  themselves  with  the  problem  of  raising  chickens  in  the 
laboratory.  They  stated,  in  accord  with  our  results,  and  in  opposi- 
tion to  those  of  Drummond,  that  it  is  quite  possible  to  raise  young 
chickens  in  cages.  On  a  diet  containing  butter  as  a  source  of  vita- 
mine  A,  and  an  addition  of  filter  paper,  it  was  possible  to  raise  some 
several  weeks  old  animals  to  normal  size.  Although  most  of  the 
animals  suffered  from  weakness  in  the  legs,  the  investigators  believed 
that  as  soon  as  all  conditions  for  the  growth  of  chickens  were  known, 
it  would  be  possible  to  conduct  growth  experiments  similar  to  those 
with  rats.  In  these  experiments,  no  green  food  was  given  and  the 
diet  was  thought  to  be  completely  carrotinoids-free,  which  was  denied 
by  Palmer.  Hart,  Halpin  and.Steenbock  (252)  were  of  the  opinion 
that  the  disease  of  chickens  mentioned  above,  designated  "leg  weak- 
ness," could  not  be  considered  as  an  avitaminosis.  Practical  chicken 
breeders  attribute  this  disease  to  the  close  confinement,  lack  of  green 
food  and  over-eating.  The  above  workers  explained  the  condition 
as  being  due  to  constipation  and  stated  that  the  green  food,  because 
of  its  vitamine  content  and -cellulose,  might  have  had  a  beneficial 
influence  on  the  constipation.  They  found  indeed  that  green  food 
is  not  necessary,  and  that  the  same  results  could  be  obtained  by  giving 
filter  paper,  dirt  or  animal  charcoal  instead;  it  was  necessary  to 
mix  these  substances  intimately  with  the  diet.  These  additions 


90  THE    VITAMINES 

served  not  only  as  roughage,  preventing  constipation,  but  also  to 
dilute  the  diet  and  make  it  more  easily  utilized.  In  most  cases,  the 
diet  consisted  of  25  per  cent  casein,  38  per  cent  dextrin,  15  per  cent 
butter,  5  per  cent  salt  mixture,  2  per  cent  agar  and  15  per  cent  dried 
yeast.  With  less  casein,  butter,  yeast  and  salts,  and  more  dextrin, 
in  making  up  a  diet  which  is  very  good  for  rats,  no  growth  was 
observed  in  chickens.  The  experiments  lasted  from  three  to  four 
months  and  it  still  remains  to  be  seen  whether  with  this  regime  the 
experiments  can  be  duplicated,  for  only  then  can  we  regard  the 
problem  of  the  synthetic  nutrition  of  chickens  as  solved.  Filter  paper 
seemed  to  be  the  best  substance  to  use  for  roughage,  although  an 
addition  of  10  per  cent  animal  charcoal  was  also  effective. 

Palmer  (253)  carried  out  a  series  of  experiments  in  which  he  tried 
to  raise  animals,  and  particularly  chickens,  on  a  diet  completely  free 
from  plant  pigments.  The  experiments  are  of  special  interest  because 
of  their  relationship  with  the  antirachitic  vitamine,  to  be  referred 
to  later.  Palmer  found  that  some  of  the  animal  pigments  belonging 
to  the  carrotinoid  group,  have  their  origin  in  the  vegetable  kingdom. 
The  species  having  a  colored  adipose  tissue  obtain  this  pigment  from 
the  blood,  in  which  it  is  easy  to  demonstrate  its  presence.  In  a  series 
of  feeding  experiments  with  chickens,  Palmer  and  Kempster  (254) 
showed  that  in  some  foodstuffs  a  certain  relationship  exists  between 
the  carrotinoid  and  the  vitamine  A  content;  they  demonstrated, 
however,  that  this  relationship  is  only  apparent  and  that  the  pig- 
ments, per  se,  play  no  part  in  metabolism.  They  also  experienced 
considerable  difficulty  in  raising  the  chickens  in  a  closed  room,  and 
further,  in  finding  a  diet  free  from  pigments.  In  the  end,  they  chose 
a  diet  composed  of  white  corn,  bran  from  the  white  corn,  bleached 
flour,  centrifuged  milk,  and  bone  meal.  The  chickens  used  were 
white  Leghorns  and  weighed  from  700  to  750  grams  at  the  start; 
the  colored  parts  of  the  body,  like  the  beak,  and  the  feather  quills, 
were  only  weakly  colored;  the  mortality  was  quite  high.  After  six 
months,  there  were  only  five  out  of  eleven  chickens  still  alive  which 
had  doubled  their  weight  and  begun  to  lay  eggs.  The  eggs  were  not 
completely  pigment-free,  but  were  nevertheless  only  weakly  colored. 
Very  young  chickens  could  not  be  raised  on  this  diet.  Better  results 
were  obtained  when  filter-paper  was  added,  according  to  the  method 
of  Osborne  and  Mendel. 


THE    VITAMINES   IN    THE    ANIMAL    KINGDOM  91 

However,  instead  of  using  butter,  which  is  not  carrotin  free,  pig 
liver  was  used  as  a  source  of  antirachitic  vitamine.  Sixty  chicks 
were  placed  on  this  diet,  upon  which  they  grew  well  for  about  6 
weeks,  but  then  began  to  lose  ground.  At  this  stage  of  the  experi- 
ment, young  pigs'  liver  was  given  with  the  result  that  an  immediate 
improvement  was  seen.  The  animals,  which  were  3  months  old,  were 
of  normal  size  and  weight  and  received  during  the  summer  an  addition 
of  pigment-free  vegetables,  such  as  white  onions.  Palmer  and 
Kempster  concluded  that  young  chickens  could  live  without  carrotin- 
oids,  if  the  diet  contained  sufficient  vifcamines.  They  believed  also 
that  the  possibility  of  discovering  the  nature  of  vitamine  A  by  an 
investigation  of  the  carrotinoids  was  without  foundation  in  fact. 
The  animals  fed  on  a  carrotinoid-free  diet  laid  eggs  after  6  months, 
and  yet  the  chickens  in  the  second  generation  were  few  in  number 
and  rather  puny  in  appeajance. 

Hart,  Halpin  and  McCollum  (255)  and  Hart,  Halpin  and  Steen- 
bock  (256)  studied  the  nutritive  requirements  of  chickens.  For  their 
experiments,  they  used  half-grown  animals  fed  on  a  mixture  of  corn, 
corn  gluten  and  calcium  carbonate,  and  also  wheat,  wheat  gluten 
and  calcium  carbonate.  The  birds  grew  well  and  were  able  to  lay 
fertile  eggs.  These  findings  stood  out  in  marked  contrast  to  the 
results  obtained  with  rats  and  pigs,  and  showed  satisfactorily  that  the 
nutritive  needs  of  chickens  and  some  mammals  were  totally  dissimilar. 
In  chickens,  an  addition  of  salts,  casein  and  butter  were  without  the 
slightest  influence,  while  the  protein  concentrates  acted  favorably. 
These  results  might  be  attributed  either  to  the  influence  of  pro- 
tein concentration  or  to  the  addition  of  unknown  factors,  which 
are  not  present  in  cereals  and  casein,  and  which  exert  a  favor- 
able influence  on  egg  production.  The  experiments  with  younger 
animals  (I.e.  256)  resulted  somewhat  differently.  In  this  case,  corn 
was  a  sufficient  source  of  B-  and  A-vitamine,  but  not  so  with  wheat, 
upon  which  the  animals  lived  only  three  months.  It  was  only  when 
salts,  casein  and  butter  were  added  that  the  diet  appeared  to  be 
complete.  In  a  similar  manner,  Harney  (257)  was  able  to  show  that 
for  the  production  of  eggs,  the  use  of  plant  foodstuffs,  in  spite  of 
the  high  protein  content,  could  not  be  compared  to  that  of  animal 
food,  and  besides  this,  Kaupp  (258)  held  that  an  addition  of  skimmed 
milk  could  influence  the  production  of  eggs  favorably. 


92  THE   VITAMINES 

In  accordance  with  these  experiments,  we  must  recognize  the 
importance  of  two  vitamines,  A  and  B,  for  the  growth  of  young 
chickens.  For  the  grown  animal,  however,  the  importance  of 
vitamine  A  had  not  been  set  forth  so  clearly.  The  recognition  of 
these  related  facts  is  important  for  the  proper  consideration  of 
chicken  beriberi,4  a  disease  arising  largely  through  feeding  with 
white  rice.  McCollum  and  Davis  (259)  were  able  to  show  that  this 
kind  of  rice  lacks,  besides  vitamine  B,  protein  of  high  biological  value, 
some  salts,  and  vitamine  A.  Whether  these  results,  obtained  with 
rats,  are  applicable  to  other  animals  without  further  work,  is  at  best 
questionable;  yet  we  must  consider  the  possibility  that  when  we  try 
to  cure  animals,  fed  with  white  rice,  by  adding  vitamine  B,  the 
indefinite  therapeutic  results  sometimes  obtained  might  be  due  par- 
tially to  the  lack  of  other  dietary  constituents. 

In  conclusion,  we  shall  mention  the  work  of  Houlbert  (260)  who 
kept  chickens  on  white  rice,  wheat  and  barley,  heated  in  an  autoclave 
to  render  them  poor  in  vitamines;  twice  a  week,  cod  liver  oil  was 
added.  After  40  days,  it  was  found  that  the  division  of  cells  in  the 
sexual  and  hematopoetic  organs  had  come  to  an  end.  However,  if 
the  missing  vitamine  was  replaced  in  the  diet,  the  glands  resumed 
normal  development. 

Chicken  beriberi.  In  relation  to  the  experiments  of  Eijkman  and 
Grijns,  chicken  beriberi,  besides  being  brought  on  by  white  rice,  may 
arise  from  feeding  white  bread  (Hill  and  Flack  261),  an  observation 
also  made  by  Ohler  (262)  and  by  Weill  and  Mouriquand  (263) .  Ohler 
also  produced  the  disease  by  feeding  hominy,  but  not  that  from  whole 
corn.  Based  upon  our  experiments,  Wellman,  Bass  and  Eustis  (264) 
used  cane  sugar  and  corn  starch;  Wellman  and  Bass  (265)  also  used 
macaroni.  Naturally,  these  experiments  are  only  of  historical  inter- 
est since  they  showed  that  the  etiology  of  beriberi  is  not  related  only 
to  the  rice  consumption.  We  have  known  for  a  long  time  that  all 
fopdjaooiLin  B-vitamine  can  give  rise  to  beriberi. 

In  the  work  of  Vedder  ancPdafk  (266),  there  is  an  excellent 
description  of  chicken  berberi.  The  first  symptoms,  on  an  exclusive 

4  In  agreement  with  some  others,  in  the  light  of  the  facts  established  in 
this  field  of  work,  I  have  dropped  the  designation  "Polyneuritis  gallinarum," 
since  according  to  the  newest  developments,  it  has  nothing  to  do  with  neuritis, 
but  perhaps  with  a  general  change  in  almost  all  of  the  tissues.  For  a  long 
time,  I  have  used  the  term  "experimental  beriberi  of  animals"  for  this 
condition. 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  93 

diet  of  white  rice,  appear  in  from  20  to  30  days.  At  first  there  is 
paralysis  of  the  extensors  of  the  legs;  the  bird  sits  on  a  flexed  tarso 
metatarsal  joint.  Paralysis  soon  extends  to  the  wings,  nape  of  the 
neck  and  the  entire  musculature.  The  animal  then  lies  motionless 
on  its  side ;  a  deep  prostration  appears  frequently  on  the  second  or 
third  day  after  the  onset  of  paralysis — at  the  latest,  in  one  week — 
and  is  followed  by  death  in  all  cases;  the  whole  course  of  the  disease 
is  run  in  a  very  short  time.  From  the  characteristic  symptoms  of 
experimental  beriberi,  the  following  should  be  noted  particularly: 

The  crest  becomes  bluish  red  very  soon,  and  this  appearance  may 
be  considered  as  one  of  the  initial  symptoms. 


FIG.  9.  AVIAN  BERIBERI 
First  day  symptoms   (Fraser-Staiiton) 

Paralysis  of  the  wing  musculature  belongs  to  the  later  symptoms — 
the  wings  then  hang  loose  down  to  the  ground.  , 

Spasticity  seldom  appears  in  the  initial  stages  of  the  disease,  but 
more  often  in  the  later  stages.  We  notice  here  the  retraction  of  the 
head  to  the  back;  sometimes  there  is  seen  the  spastic  walk  on  tip- 
toes with  stiff  knees.  Dysphagia  belongs  to  the  early  symptoms; 
water  given  to  the  bird  flows  out  again  from  the  beak;  on  attempting 
to  feed  forcibly,  the  pigeon  chokes.  According  to  Vedder  and  Clark, 
the  loss  of  weight  is  constant  at  about  20  per  cent  of  the  initial  weight. 

Progress  of  the  disease.  The  above  named  investigators  describe 
the  progress  of  the  disease  in  the-  following  manner :  In  acute  cases, 
with  prostration  and  great  loss  of  weight,  all  the  symptoms  are 


94  THE    VITAMINES' 

apparent  in  24  hours.  The  bird  lies  on  one  side,  shows  dyspnoea  and 
cyanosis  and  dies  in  a  few  hours.  The  second  type  is  more  chronic; 
in  these  cases  we  see  paralysis  of  the  legs,  but  good  general  condition. 
The  appetite  remains  good,  the  loss  in  weight  is  insignificant;  the 


FIG.  10.  AVIAN  BERIBERI 
First  day  symptoms  (Fraser-Stanton) 

crest  remains  red ;  in  this  condition,  the  chicken  may  live  for  weeks. 
According  to  our  personal  experience,  the  second  type  is  not  so  suit- 
able for  curative  experiments. 

Segawa  (267)  also  described  two  types  of  beriberi — one  form  which 
he  regarded  as  a  pure  polyneuritis,  and  another,  possessing  more  of 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOMS 


95 


the  characteristics  of  starvation,  and  expressed  by  a  distinct  aversion 
to  rice.  In  66  per  cent  of  all  cases,  both  types  occurred  together. 
In  Segawa's  experiments,  some  animals  remained  in  good  condition 
for  about  219  days;  this  may  perhaps  have  been  due  to  the  possibility 
that  the  animals  had  eaten  traces  of  other  food,  such  as  flies  and 
vermin.  He  erroneously  considered  the  disease  as  a  real  polyneuritis. 
Tasawa  (268)  observed  that  the  marked  emaciation  and  starvation 
could  be  avoided  by  an  addition  of  egg-yolk  or  cooked  meat,  where- 
upon only  clear  beriberi  symptoms  are  noted. 

As  we  shall  see  later  in  pigeons,  fatigue  plays  a  big  part  in  the 
development  of  acute  symptoms.     In  accord  with  this,  Hulshoff  Pol 


FIG.  11.  AVIAN  BERIBERI 
Second  day  symptoms  (Fraser-Stanton) 

(269)  found  that  forced  feeding  accelerates  the  acute  symptoms,  and 
Williams  and  Johnston  (270)  found  that  the  same  effect  could  be 
hastened  by  increased  temperature  and  exercise. 

Pigeons 

The  problem  as  to  the  normal  food  requirements  of  pigeons  appears 
to  be  much  simpler  than  that  of  chickens.  Since  pigeons  may  be 
raised  in  the  laboratory  with  little  difficulty  and  since  they  do  not 
eat  vermin  and  flies,  the  feeding  experiments  may  be  conducted  with 
greater  exactness.  Young  pigeons,  after  they  have  been  hatched,  are 
fed  alternately  by  their  parents,  so  that  it  is  possible,  through  the 


96  THE   VITAMINES 

administration  of  a  special  diet  to  the  latter,  to  study  its  effect  upon 
the  young  from  the  very  beginning. 

We  have  personally  kept  grown  pigeons  on  an  exclusive  corn  diet 
for  more  than  a  year,  during  whch  time  they  were  not  only  in  splendid 
health  but  reproduced  normally.5  It  cannot  be  emphasized  suffi- 
ciently, that  whole  corn  provides  sufficient  nutrition  for  the  maintenance 
and  growth  of  pigeons,  which  indicates  that  corn  is  quite  sufficient  for 
at  least  one  species.  This  is  of  particular  importance  for  our  later 
conception  of  the  etiology  of  pellagra,  which  has  recently  been  attrib- 
uted to  the  deficiency  of  corn  proteins.  That  corn  is  a  complete 
food  for  pigeons  has  been  shown  by  other  workers.  Voegtlin  and 
Myers  (271),  reported  that  they  maintained  pigeons  in  good  health  on 
an  exclusive  diet  of  corn  for  at  least  four  months.  In  another  series 
of  experiments,  the  same  workers  (272)  investigated  the  conditions 
of  growth  in  young  pigeons  which  attained  normal  size  in  40  days, 
after  having  been  fed  by  their  parents  who  had  been  given  a  diet  of 
whole  corn  or  whole  wheat,  with  the  addition  of  a  calcium  salt.  The 
same  result  was  obtained  when  the  diet  was  composed  of  white  bread, 
with  the  addition  of  vitamine  B  and  calcium  salts;  and  also  wheat 
starch,  casein,  A-  and  B-vitamine,  and  calcium  salts.  Because  of  the 
possible  interest  in  the  study  of  the  growth  of  young  pigeons,  there  is 
appended  a  diagram  illustrating  normal  growth  as  compared  with 
growth  on  corn,  taken  from  the  work  of  Voegtlin  and  Myers. 

It  is  shown  that  young  pigeons  require  two  kinds  of  vitamines, 
namely,  A-  and  B-vitamines,  and  can  get  along  very  well  in  the 
entire  absence  of  C-vitamine,  which  is  in  direct  opposition  to  the 
needs  of  most  mammals.  Regarding  the  requirements  of  grown  pigeons 
for  vitamine  A,  this  must  be  left  for  the  future  to  determine.  Should 
it  appear  that  adult  pigeons  also  need  vitamine  A,  it  would  be  worth* 
while  to  so  change  the  usual  diet  used  to  demonstrate  the  onset  of 

6  Among  these  were  also  pigeons  which  after  being  cured  of  beriberi,  still 
showed  normal  fecundity. 

6  Experiments  were  made  by  us  (273),  in  which  two  pigeons  lived  for  49 
to  54  days  respectively  on  an  artificial  diet  if  B-vitamine  was  injected  every 
few  days,  and  finally  died  of  sepsis  caused  by  contamination  of  the  vitamine 
solution.  Later  on,  Stepp  (274)  carried  out  some  similar  experiments  in 
which  he  kept  a  pigeon  in  excellent  health  for  91  days  on  dog  biscuits  extracted 
with  alcohol,  and  rice  polishings  (orypan) ;  when  the  orypan  was  discontinued, 
the  bird  died  in  37  days.  In  spite  of  the  use  of  orypan,  the  bird  could  not 
fly  well  and  lost  some  weight. 


THE   VITAMINES   IN   THE    ANIMAL   KINGDOM 


97 


beriberi,  namely,  white  rice,  that  it  would  lack  nothing  but  B-vita- 
mine,  although  we  were  able  to  induce  a  typical  beriberi  with  a  syn- 
thetic diet  lacking  vitamine  A  (275) .  As  we  have  already  pointed  out 
in  the  case  of  the  chicken,  all  these  dietary  deficiencies  can  complicate 
the  picture  of  experimental  beriberi  unnecessarily.  Because  of  the 


FIG.  12.  GROWTH  CURVES  OF  SQUABS 

Parents  were  fed  on  a  diet  limited  to  the  corn  and  wheat  kernel,  supple- 
mented by  the  addition  of  calcium  carbonate  (oyster  shells)  (Voegtlin-Myers), 

protein  deficiency  of  white  rice,  Dutcher  (276),  and  others,  used  an 
addition  of  3  per  cent  casein,  calculated  on  the  dry  weight  of  rice. 

Pigeon  beriberi.  The  method  most  commonly  used  to  induce  beri- 
beri in  pigeons  is  to  feed  white  rice,  although  other  foodstuffs  and 
mixtures  of  isolated  substances  are  available  for  that  purpose. 


98  THE    VITAMINES 

Among  others,  Funk  and  Cooper  (277)  showed  in  1911  that  the  dis- 
ease could  be  brought  about  with  pure  sugar,  inulin,  dextrin  and 
starch.  The  pigeon  is  quite  the  best  animal  to  use  in  the  study  of 
beriberi.  When  pigeons  are  placed  upon  a  diet  of  polished  rice, 
they  eat  it  with  great  avidity  during  the  first  few  days.  Following 
this,  their  appetite  diminishes  markedly,  and  they  frequently  attempt 
to  disgorge  the  rice  out  of  their  crops.  This  behavior  is  more  often 
noticed  in  animals  that  are  fed  forcibly,  but  sometimes  this  rice  is 
again  eaten  voluntarily.  This  condition  makes  it  difficulty  to  carry 
out  exact  rice  feeding  experiments  with  pigeons.  One  has  the  impres- 
sion that  the  animals  have  an  aversion  for  rice,  which,  however,  dis- 
appears if  the  necessary  vitamine  is  administered  at  the  same  time 
with  the  food.  After  a  few  days,  it  is  best  to  put  the  pigeon  in  a  box 
with  a  wooden  cover  so  that  its  head  extends ;  in  this  way  it  is  possible 
for  one  person  to  manage  when  forcible  feeding  must  be  resorted  to. 
The  beak  is  opened  and  through  a  small  metallic  funnel  with  smooth 
edges,  the  weighed  rice  is  pushed  into  the  crop  with  a  glass  rod. 
Another  method  used  by  us  was  to  prepare  the  food  in  the  form  of 
pills,  and  feed  them  as  such.  For  feeding  purposes,  it  is  best  to  use 
healthy  males  weighing  from  300  to  350  grams;  20  to  30  grams,  of 
rice  per  day  may  be  given  in  three  portions.  If  the  crop  should 
happen  to  be  full,  it  is  necessary  to  wait  till  it  is  empty.  We  shall 
speak  of  the  effect  of  the  food  ingested  when  we  come  to  the  physi- 
ology of  vdtamine  B  (p.  210).  After  several  days  of  rice  feeding,  a 
marked  change  is  noted  in  the  animals.  Very  little  remains  of  the 
usually  predominating  desire  to  quarrel  when  a  number  of  pigeons 
are  kept  in  one  cage.  Likewise,  the  sexual  instinct,  such  as  the 
strutting  of  the  male  around  the  female,  seems  to  be  held  in  abeyance, 
and  the  animals  sit  on  the  perch  in  a  sleepy,  apathetic  manner.  They 
experience  increasing  difficulty  in  flying  on  to  the  perch  and  a  few 
days  later  some  of  the  pigeons  sit  on  the  bottom  of  the  cage,  regardless 
of  the  fact  that  they  are  soiled  by  the  feces  of  the  animals  on  the 
perch  above.  The  fecal  matter,  normally  of  semi-solid  consistency 
and  whitish  color,  becomes  slimy  and  water-clear,  or  slight  yellowish, 
and  the  animals  show  no  disposition  to  keep  themselves  clean.  If 
they  are  left  to  feed  themselves,  their  appetite  disappears  gradually 
and  their  condition  is  that  of  semi-starvation.  About  30  per  cent  of 
the  animals  which  are  permitted  to  feed  themselves  develop  symp- 
toms of  beriberi,  while  the  remainder  die  of  general  weakness.  Of 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM  99 

the  animals  forcibly  fed,  a  greater  proportion  develop  the  typical 
beriberi  symptoms,  after  a  period  of  time  which  is  subject  to  great 
deviations.  We  may  say,  however,  that  most  of  the  animals  develop 
the  disease  in  from  10  to  30  days;  it  is  noticeable  that  the  animals 
mostly  become  sick  within  a  few  days  of  each  other,  so  that  there  is 
the  false  impression  of  the  appearance  of  an  epidemic.  In  the 
individual  animals,  we  note  the  development  of  various'  types  of 
diseases,  as  illustrated  in  the  accompanying  pictures,  closely  resem- 
bling the  conditions  described  in  man.  This  analogy  may  well  be 
fallacious  and  the  types  described  below  may  depend  perhaps  only 
on  the  nutritive  condition  and  the  length  of  time  the  rice  is  fed. 

In  one  type,  which  we  shall  designate  as  acute,  we  notice  that  the 
ability  to  walk  is  decreased.  If  the  animal  is  frightened,  it  runs  a 
few  steps  without  difficulty,  but  when  fatigue  sets  in,  the  animal 
helps  itself  by  flapping  its  wings,  but  sits  motionless  when  left  alone; 
the  heart  beat  is  pronounced  and  behaves  as  though  it  had  been  over- 
exerted. The  next  symptom  is  uncontrolled  motions  of  the  head. 
If  the  animal  is  turned  about  several  times  in  the  air,  then  in  many 
cases  the  acute  form  is  produced  out  of  the  latent.  The  phenomenon 
appears  quite  suddenly — the  head  is  pressed  against  the  back  by  a 
retraction  of  the  neck  muscle,  the  legs  are  drawn  up  to  the  belly; 
the  animal  turns  somersaults,  and  this  may  last  a  long  time  under 
certain  conditions.  In  this  stage,  the  animal  does  not  survive  very 
long.  During  the  course  of  a  day,  a  disturbance  in  breathing 
develops;  the  animal  opens  its  beak  wide,  apparently  struggling 
against  suffocation,  and  finally  death  results  in  from  12  to  24  hours 
after  the  appearance  of  the  symptoms,  with  an  easing  up  of  the 
spasms. 

The  second  or  chronic  type  (figs.  14-15)  develops  as  follows: 
After  a  few  weeks  the  animal  is  found  sitting  in  the  cage;  it  moves 
very  reluctantly  in  spite  of  its  ability  to  do  so.  After  some  days, 
it  loses  even  this  capability,  and  sits  motionless  in  one  spot.  Only 
seldom  does  this  form  go  over  into  the  spastic.  Usually,  the  animal 
lives  a  few  weeks  longer  and  dies  without  moving  from  its  place. 
These  cases  are  not  suitable  for  curative  experiments.  For  this 
purpose,  we  must  use  only  those  animals  in  the  first  group. 

The  latter  also  show  variations.  In  some,  the  spastic  condition 
persists  till  death  while  in  others,  it  disappears  for  short  intervals 
only  to  reappear  in  a  more  severe  form. 


100 


THE   VITAMINES 


FIG.    13.  SPASTIC  FORM  OF  BERIBERI  IN  PIGEONS 


FIG.  14.  ATROPHIC  FORM 


THE    VITAMINES   IN   f^E 'ANIMAL 


101 


FIG.  15.  PARALYTIC  FORM 


FIG.  16.  SPASTIC-PARALYTIC  FORM 


102 


THE'  VIT^MINES 


A  further  phenomenon,  common  to  both  forms  of  beriberi  in 
pigeons,  is  the  loss  in  weight,  which  is  apparent  before  the  onset  of 
the  symptoms.  This  loss,  amounting  to  from  20  to  45  per  cent  of  the 
original  weight,  occurs  always  in  pigeons  fed  on  rice,  according  to  our 
experience.  The  loss  in  weight  is  apparent  whether  the  animal  is 
fed  forcibly  or  not.  Results  to  the  contrary  are  due  to  the  fact  that 
the  pigeons,  chiefly  because  of  the  paralysis  of  the  muscles  of  the 
crop,  are  unable  to  empty  it.  On  post  mortem,  the  crop  is  tightly 
packed  with  rice,  weighing  about  100  grams,  which  ordinarily  would 
have  been  calculated  as  body  weight.  It  cannot  be  emphasized 
sufficiently  that  in  experimental  beriberi,  following  a  strictly  managed 


400 

350 
300 
250 
200 
IPO 
100 

0          *          8         K         16       U 

\ 

—  __ 

\ 

^ 

X 

\J 

^ 

\ 

^^ 

^ 

\ 
\ 

\ 

FIG.  17.  DROP  IN  WEIGHT  OF  PIGEONS  FED  ON  30  GRAMS  POLISHED  RICE  DAILY 

diet,  death  ensues  in  100  per  cent  of  the  cases.  Theiler,  Green  and 
Viljoen  (278)  have  observed  that  starving  pigeons,  only  on  water, 
may  show  spontaneous  cures.  This  behavior  was  explained  by  the 
mobilization  of  the  vitamine  in  the  tissues.  The  tissues  are  catabo- 
lized  more  rapidly  during  starvation  and  therefore  a  greater  supply 
of  vitamine  may  suddenly  be  made  available  for  the  animal.  If  rice 
feeding  is  continued  death  must  eventually  ensue.  The  question  of 
malnutrition  in  beriberi  was  also  taken  up  by  Lumiere  (279). 

Besides  chickens  and  pigeons,  beriberi  has  also  been  described  in 
other  birds,  which  might  sometimes  also  be  of  practical  importance. 
For  instance,  Merklen  (280)  described  a  disease  in  3  or  4  weeks  old 
ducks  manifesting  itself  in  symptoms  of  cramps,  paralysis  of  the  legs 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM  103 

and  inanition,  which  quickly  disappeared  on  substituting  the  diet 
by  one  more  varied.  Perhaps  we  are  dealing  with  beriberi  here 
too.  The  appearance  of  beriberi  in  other  birds  is  tabulated  below: 

Animal  Observer 

Ducks Eijkman  (I.e. 48)  and  Ktilz  (281) 

Geese Eijkman  (l.c.48) 

Sparrows Fujitani  (282) 

Quail Toyama  (283) 

Jushimatsu Toyama  (283) 

Parrots Fink  (284) 

Rice  birds Ottow  (518) 

Munia  maja Jansen  (284a) 

Pathological  anatomy  and  chemical  pathology  of  beriberi  in  birds 

Pathological  anatomy.  Till  recently,  everyone  was  of  the  opinion 
that  in  avian  beriberi  the  manifestations  in  the  central  nervous 
system  were  in  the  foreground.  The  new  investigations  show,  on 
the  other  hand,  that  as  a  result  of  the  lack  of  vitamine  B,  many,  if 
not  all,  of  the  organs  suffer,  and  for  this  reason  the  disease  cannot  be 
thought  of  as  "polyneuritis."  Despite  many  attempts,  it  has  not  yet 
been  possible  to  find  out  which  organ  or  tissue  is  primarily  responsible 
for  the  manifestations  of  the  disease.  This  is  of  course  bound  up  with 
our  insufficient  knowledge  of  the  physiology  and  significance  of  the 
vit  amines. 

Nervous  system.  We  find  a  very  good  description  of  it  in  the 
work  of  Vedder  and  Clark  (I.e.  266),  already  mentioned,  and 
more  recently  in  a  report  by  Onari  Kimura  (285).  We  see  from 
these  investigations,  that  not  .all  nerves  undergo  pathological 
changes  in  the  same  measure;  for  example,  the  vagus  degen- 
erates, but  not  in  a  high  degree.  However,  in  the  opinion  of 
Kimura,  beriberi  is  a  general  disease  of  the  nervous  system,  and 
this  was  our  opinion  too.  We  still  believe  that  all  of  the  hetero- 
geneous pathological  changes  .occurring  in  beriberi  may  best  be 
looked  upon  as  of  central  origin,  in  so  far  as  they  may  all  be  regarded 
as  atrophic  changes.  Naturally,  we  admit  that  still  another  inter- 
pretation may  be  found. 

The  nerves  of  the  lower  extremities  are  selectively  more  affected 
than  those  of  the  upper.  Especially  is  this  true  of  the  sciatic  and 
peroneus  nerves  which  show  a  fatty  degeneration,  though  no  definite 


104  THE   VITAMINES 

paralysis  is  noted.  These  changes  arise  after  7  days  of  rice  feeding. 
The  number  of  degenerated  fibers  bears  no  relation  to  the  severity 
of  the  paralysis.  Cases  of  light  clinical  symptoms  often  display 
marked  degeneration,  while  severe  cases  frequently  show  only  from 
4  to  10  per  cent  of  degenerated  fibers.  As  a  rule,  from  10  to  15  per 
cent  of  the  total  fibers  are  found  changed.  Schnyder  (286)  on  the 
contrary,  finds  only  slight  degeneration  in  birds  and  believes, 
because  of  the  therapeutic  influence  of  the  vitamine,  that  the  paresis 
could  not  be  the  result  of  degenerative  processes  in  the  brain.  We 
have  frequently  pointed  out  that  if  we  kill  an  animal  a  few  days 
after  it  has  been  cured  with  vitamine,  and  make  a  histological  study 
of  the  nerves,  the  degenerated  fibers  are  still  to  be  seen  and  persist 
for  a  long  time.  In  the  meantime,  the  normal  fibers  appear  to 
assume  the  functions  of  the  diseased  fibers. 

The  nerves,  histologically  examined,  give  the  following  picture: 
According  to  Kimura,  the  first  indication  of  degeneration  is  to  be 
sought  for  in  the  axis  cylinder,  while  the  earlier  prevalent  idea  was 
that  the  signs  of  degeneration  are  first  noted  in  the  medullary  sheath; 
the  myelin  sheath,  according  to  Kimura,  may  remain  intact.  How- 
ever, as  soon  as  the  latter  degenerates,  the  axis  cylinder  can  no  longer 
be  differentiated.  The  myelin  fragments  thereupon  are  resorbed 
in  situ  through  the  cells  of  Schwann's  sheath.  If  a  degenerated 
myelin  fiber  still  contains  an  axis  cylinder,  it  is  a  regenerated  axis 
cylinder  from  the  ribbon-like  protoplasm.  The  new  cylinder  is 
smooth  and  fragile  and  resembles  that  seen  after  a  trauma. 

The  same  changes  are  also  to  be  noted  in  the  dorsal  and  ventral 
nerve  roots,  as  in  all  the  dorsal  strands  of  the  spinal  cord.  Further, 
changes  are  apparent  in  the  cells  of  the  forward  and  rear  horns  of  the 
lumbo-sacral  cord;  here  the  tigroid  bodies  are  invisible,  and  the 
stainable  substance  (NissFs  method)  accumulates  over  the  axis 
cylinder.  'In  some  cases,  the  nuclei  are  weakly  colored.  Similar 
pathological  nerve  studies  were  made  by  Weill  and  Mouriquand  (287), 
Kato  and  Shizume  (288)  and  Paguchi  (289). 

The  muscles.  The  muscles  exhibit  atrophy  and  fatty  degeneration, 
but  the  changes  disappear  rapidly  on  returning  to  normal  nutrition. 

The  heart.  In  most  cases,  this  is  unchanged ;  only  seldom  is  there 
any  edema,  pigmentation,  and  traces  of  parenchymatous  degenera- 
tion. Hypertrophy  of  the  right  heart,  as  in  human  beriberi,  does  not 
take  place  in  chickens.  On  the  contrary,  we  have  personally  seen 


THE    VITAMINES   IN   THE    ANIMAL   KINGDOM  105 

hydropericardium   in    pigeons.    McCarrison    (290)    attributes   the 
edema  to  the  enlargement  of  the  suprarenals, 

Endocrine  glands.  The  first  investigations  in  this  direction  were 
made  by  Funk  and  Douglas  (291).  The  findings  in  pigeons  showed 
that  the  thymus  glands  disappear;  this  was  also  noted  by  R.  R. 
Williams  and  Crowell  (292).  The  behaviour  of  this  gland  in  birds 
was  then  investigated  further  by  McCarrison  (293).  Funk  and 
Douglas  made  a  histological  investigation  of  pituitary,  adrenals, 


FIG.  18.  FATTY  DEGENERATION  OF  SCIATIC  NERVE  IN  CHICKEN  BERIBERI 

ovary,  testicle,  kidney,  liver,  pancreas  and  spleen.  In  all  these 
organs,  definite  signs  of  degeneration  were  noted.  The  investigation 
of  thyroid  in  greater  detail  was  made  by  Douglas  (294)  who  observed 
carefully  the  great  variations  and  the  tendency  of  the  colloids  to 
disappear  from  the  vesicles.  McCarrison  (295)  studied  the  influence 
on  the  thyroid  of  food  poor  in  vitamine,  and  found  that  the  size  of  the 
gland,  as  well  as  the  weight,  decreased.  The  study  of  the  endocrine 
glands  of  pigeons  was  then  undertaken  by  the  same  worker  (296) 
who  largely  corroborated  the  work  of  Funk  and  Douglas,  and  found 


106  THE   VITAMINES 

that  the  organs  undergo  atrophy  in  the  following  order:  thymus, 
testicle,  spleen,  ovary,  pancreas,  heart,  liver,  kidney,  stomach, 
thyroid  and  brain.  McCarrison  made  the  very  interesting  and 
important  observation  that  the  suprarenals  axe  often  hypertrophied,7 
and  that  this  hypertrophy  bore  a  causative  relationship  to  the  onset 
of  edema  (I.e.  290).  He  believed  that  in  unpolished  rice,  butter 
(particularly  in  butter,  as  will  be  noted  later)  and  onions,  there  was  a 
substance  which  was  protective  against  edema,  and  which  might  be 
thought  of  as  vitamine  A. 

McCarrison  found  also  that  the  brain,  suprarenals  and  the  pitui- 
tary, are  very  sensitive  to  a  lack  of  vitamine.  The  sexual  organs 
in  males  showed  a  disappearance  of  spermatogenesis,  with  resulting 
sterility;  in  females,  there  was  a  condition  similar  to  amenorrhea. 
This  is  in  opposition  to  the  fact  that  we  mated  pigeons,  cured  of  beri- 
beri, and  secured  a  completely  normal  progeny.  It  is  nevertheless 
possible  that  in  these  cases,  there  may  have  been  a  regeneration. 

During  the  course  of  his  fruitful  investigations,  McCarrison  also 
examined  the  muscles  and  found  them  highly  atrophied,  while  the 
central  nervous  system  was  only  slightly  so;  he  attributed  the  result- 
ant paralytic  symptoms  directly  to  the  impaired  functional  ability 
of  the  nerve  cells.  Because  of  the  remarkable  atrophy  of  the  thymus  r 
testicles,  ovaries,  and  spleen,  much  more  apparent  than  in  other 
organs,  resulting  from  the  lack  of  vitamines,  he  believed  that  these 
organs  provided  the  distressed  organism  with  the  needed  vita- 
mine.  When  this  reserve  is  exhausted  then  the  bones  are  the 
purveyors  of  the  vitamine,  in  which  case  the  marrow  undergoes 
marked  changes.  Red  corpuscles  are  also  diminished  about  25  per 
cent.  Finally,  McCarrison  regarded  the  whole  picture  of  beriberi  as 
a  syndrome,  arising  from,  (1)  a  chronic  inanition;  (2)  a  pathological 
change  of  the  organs  of  digestion  and  assimilation;  (3)  an  abnormal 
functioning  of  the  internal  secretory  glands,  especially  of  the  supra- 
renals; and  (4)  poor  nutrition  of  the  central  nervous  system.  In 
addition  to  all  these  factors,  there  is  also  a  decreased  resistance  to 
the  bacterial  invasion.  McCarrison  (297)  at  first  erroneously 
attributed  to  the  latter  condition  the  entire  picture  of  beriberi,  but 
he  has  long  since  abandoned  this  view.  The  pathological  histology 
of  beriberi  in  pigeons  is  described  by  McCarrison  (298)  in  a  special 
article. 

7  Unfortunately,    in  the  pathological   department,   which  took    up    this 
phase  of  the  work,  this  observation  completely  escaped  notice. 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM 


107 


Changes  in  the  g astro-intestinal  tract.  McCarrison's  (299)  investiga- 
tion on  153  pigeons  yielded  some  very  interesting  results.  The 
upper  part  of  the  intestine,  which  is  muscular  in  nature  in  the  region 
of  the  pancreas,  is  mostly  involved.  Atrophy  had  proceeded  to 
such  an  extent  that  the  walls  were  almost  transparent,  especially 
when  white  rice  was  fed  for  a  long  time.  The  intestine  showed  a 


FIG.  19.  THE    UPPER    PART    OF    THE    INTESTINE    OF    A    HEALTHY    PIGEON 

(McCARRISON) 

definite  congestion;  ecchymosis  was  not  infrequently  seen  and 
microscopic  hemorrhages  were  almost  always  observed.  Under  the 
microscope,  the  congestion  led  to  a  partial  disappearance  of  the 
epithelium.  The  myenteron  was  so  severely  atrophied,  that  its 
motor  functions  suffered.  Further,  there  was  described  degeneration 
of  the  mesenteric  plexus  of  Auerbach,  which  impaired  the  functions 


108 


THE    V1TAMINES 


of  the  intestinal  nerves.  The  villi  disappeared  and  there  was  atrophy 
and  inflammation  of  the  mucosa  and  atrophy  of  the  lymphatic  tissue. 
The  latter  condition  is  primarily  responsible  for  the  decreased 
resistance  to  infection.  According  to  McCarrison  (300),  the  severe 
intestinal  infection  is  the  chief  reason  whv  some  animals  cannot  be 


FIG.  20.  SECTION  OF  UPPER  PART  OF  INTESTINE  FROM  A  PIGEON  DYING  OP 

BERIBERI 

Thinning  of  muscular  coat,  intense  atrophy  and  necrosis  of  the  mucous 
membrane. 

saved  by  an  addition  of  vitamine.  Pigeons  fed  on  autoclaved  rice, 
butter  and  onions  develop  the  same  pathological  picture,  with  the 
difference  that  the  changes  are  not  so  far  advanced.  After  the 
elimination  of  butter,  it  may  be  shown  that  onions  have  the  power 
to  protect  the  intestinal  canal  against  degeneration.  By  the 
addition  of  peas,  containing  vitamine,  the  pigeons  may  be  cured, 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  109 

while  the  pathological  picture  of  the  intestine  remains  the  same.  In 
his  opinion,  the  cure  is  so  rapid  that  it  cannot  be  due  to  the  vitamine 
absorption;  the  therapeutic  effect  may  be  compared,  in  its  velocity, 
only  to  the  sudden  manifestation  of  the  growth  impulse  in  germinat- 
ing seeds. 

Chemical  pathology.  The  chemical  investigations  are  not  yet  so 
numerous.  Schaumann  (I.e.  2)  investigated  the  brains  of  a  beriberi- 
pigeon  and  found  that  the  phosphorus  content  was  not  decreased. 
In  contrast  to  this,  we  (301)  found  that  the  nitrogen  and  phosphorus 
content  of  these  brains  is  below  the  normal  content;  indeed  in 
normal  pigeons,  we  found  9.77  per  cent  nitrogen  and  1.84  per  cent 
phosphorus,  while  in  beriberi  pigeons,  there  was  9.31  per  cent  nitrogen 
and  1.53  per  cent  phosphorus.  Our  results  were  confirmed  byWie- 
land  (302)  and  by  Mathilde  Koch  and  Voegtlin  (303),  although  it  is 
not  certain  just  how  ordinary  starvation  would  affect  these  figures. 

Funk  and  v.  Schonborn  (304)  also  showed  that  in  pigeon-beriberi, 
there  is  an  increase  in  blood  sugar  and  a  decrease  in  the  glycogen 
content  of  the  liver  almost  to  the  point  of  complete  disappearance. 
Because  of  this,  we  were  very  much  inclined  to  bring  these  facts 
into  causative  relationship  with  the  observation  of  McCarrison  on 
hypertrophy  of  the  adrenals. 

It  must  also  be  recorded  that  Fuji  (305)  found  the  diastase  content 
of  the  blood  decreased  in  chickens  suffering  from  beriberi ;  on  admin- 
istration of  vitamine,  this  value  returned  to  normal.  Analogous 
results  were  obtained  with  glyoxalase  of  the  liver  by  Findlay  (305a). 

MAMMALS 

In  1913,  we  stated  that  certain  animals  do  not  develop  beriberi 
when  they  are  put  on  a  diet  of  white  rice;  we  attempted  to  explain 
this  by  its  dependence  on  the  specific  purine  metabolism  of  the 
particular  species.  With  this  in  view,  we  classified  mammals  into 
two  broad  divisions,  one  in  which  the  end  product  of  purine  metabo- 
lism is  uric  acid,  and  the  other  in  which  it  is  allantoine.  The  first 
division  included  man  and  birds,  manifesting  real  beriberi,  while  the 
second  included  the  rat,  the  dog,8  the  monkey — which  can  contract 

8  Bearing  upon  this,  it  would  be  interesting  to  determine  if  a  different 
reaction  to  the  lack  of  B-vitamine  is  manifested  by  Dalmatian  dogs,  which, 
according  to  S.  R.  Benedict  (307),  excrete  uric  acid  chiefly  instead  of  allan- 
toine, and  by  anthropoid  apes,  found  by  Hunter  and  Ward  (308)  to  have  a 
purine  metabolism  similar  to  that  of  man. 


110  THE    VITAMINES 

scurvy — while  guinea  pigs  with  their  characteristic  purine  metabo- 
lism occupy  a  middle  position.  Whether  these  conceptions  may 
still  be  regarded  as  correct  is  not  certain,  for  in  the  first  place,  a 
type  of  beriberi  in  dogs  and  rats  has  been  described  and  secondly, 
there  is  a  possibility  of  another  explanation  of  the  above  mentioned 
observations.  At  all  events,  we  wish  to  leave  the  question  open  for 
further  investigation,  particularly  since  beriberi  has  not  been  defi- 
nitely demonstrated  in  certain  animals,  and  may  even  be  confused 
with  other  pathological  conditions.  The  criteria  for  the  recognition 
of  beriberi,  such  as  the  fatty  degeneration  of  the  nerves,  are  not 
specific  symptoms  of  beriberi.  We  have  fed  white  rice  for  along 
time  to  rats,  rabbits  and  guinea  pigs,  without  noticing  any  kind  of  a 
symptom  to  suggest  beriberi.  The  other  explanation,  of  the  differ- 
ent behavior  of  certain  animals  on  a  diet  of  white  rice,  we  find  in 
the  unknown  role  of  the  bacterial  intestinal  flora,  the  significance  of 
which  has  already  been  mentioned  (p.  76)  in  the  chapter,  "Life 
Without  Bacteria."  We  come  now  to  the  consideration  of  the  impor- 
tance of  the  vitamines  for  various  mammals. 

Rats 

Rats  have  been  extensively  used  as  experimental  animals  in  the 
last  10  years  in  the  attempt  to  solve  the  problems  of  nutrition,  and 
for  vitamine  research.  The  reason  for  this  is  that  rats  breed  easily, 
do  not  consume  much  food  and  are  not  particular  in  their  taste.  The 
fact  that  a  greater  number  of  these  animals  may  be  used  at  one  time 
for  an  investigation  makes  the  results  much  more  certain.  Since  in 
most  problems  of  nutrition,  the  food  must  be  specially  prepared,  as 
a  rule,  the  small  amount  ingested  is  of  practical  significance.  Rats 
live  about  3  years  and  reach  sexual  maturity  very  early.  The 
pregnancy  lasts  3  weeks  and  the  offspring  is  numerous;  the  number 
of  young  varies  from  6  to  12  according  to  the  age  of  the  parents  and 
their  nutritive  condition.  The  young  rats  begin  to  eat  of  their  own 
accord  after  3  to  4  weeks,  and  may  be  used  for  feeding  experiments 
when  they  weigh  30  grams,  though  it  is  better  to  wait  till  they 
weigh  from  40  to  60  grams.  It  has  been  shown  by  many  investiga- 
tors, among  them  McCollurn,  Simmonds  and  Pitz  (309),  that  inade- 
quacies of  the  diet  are  manifested  in  a  diminished  milk  secretion  of 
the  mother,  or  else  in  a  poor  state  of  health  of  the  young,  although 
'  the  parents  show  no  such  signs.  It  is  therefore  desirable,  if  one 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM 


111 


wishes  to  demonstrate  the  nutritive  value  of  a  certain  diet  to  carry 
out  feeding  experiments  with  the  same  material  on  the  next  genera- 
tion. For  such  a  purpose,  the  rat  is  naturally  specially  suited. 
Thus,  it  is  possible  to  study,  in  a  short  time,  not  only  the  effect  of 
the  diet  upon  the  parents,  but  also  to  extend  the  observations  on  the 
procreative  power,  the  number  of  living  young,  and  the  lactation  of 
the  mother.  Judging  by  our  experience,  we  deem  it  inadvisable  to 


FIG.  21.  RAT  CAGE  (MACALLUM) 

6,  water  cup;  7,  flask  for  urine  collection;  8,  bulb  by  means  of  which  urine, 
is  separated  from  feces  (urine  flows  on  the  outside  of  the  bulb  into  the  flask 
and  feces  falls  into  beaker) ;  9,  food  cups  of  various  sizes;  10,  funnels  of  various 
sizes  adapted  to  the  age  of  the  animal,  so  that  the  food  may  not  be  scattered. 

buy  rats  for  experimental  purposes;  it  is  far  better  to  raise  them  in 
the  laboratory.  In  this  way  we  have  the  advantage  of  knowing  the 
parentage  of  the  animals  to  be  used.  When  one  obtains  a  litter,  it  is 
sometimes  difficult  to  differentiate  between  the  male  and  female. 
To  obviate  this,  we  can  utilize  a  method  described  by  Jackson  (310). 
It  is  likewise  of  importance  to  use  a  cage  so  arranged  as  to  be  able  to 
control  accurately  the  amount  of  food  given,  especially  since  rats 


112  THE   VITAMINES 

have  a  tendency  to  scatter  their  food.  This  difficulty  may  be  over- 
come if  we  use  such  a  cage  as  has  been  described  explicitly  by  our 
former  co-worker,  A.  B.  Macallum  (311).  The  cage  is  equipped 
with  funnels  of  various  sizes  through  which  the  rat  may  reach  its 
food  without  scattering  it.  The  cage  is  arranged  for  two  rats  and 
permits  of  separating  the  urine  from  the  feces,  so  that  an  exact  metabo- 
lism experiment  may  be  carried  out.9  Great  care  is  necessary  in  the 
preparation  of  the  diet;  above  all  it  is  important  that  the  basal 
diet  is  vitamine-free.  This  precaution  was  not  sufficiently  observed 
in  the  older  investigations,  thus  giving  rise  to  differences  of  opinion. 
We  need  only  quote  one  example,  lactose,  widely  used  as  a  constituent 
of  an  artificial  diet.  Regarding  this,  we  showed10  that  lactose,  which 
does  not  undergo  any  particular  purification  process,  contains  traces 
of  nitrogen.  These  traces  may  come  from  the  milk  and  permit  of 
the  growth  of  rats,  because  of  its  vitamine  content.  This  was 
demonstrated  experimentally  by  McCollum  and  Davis  (I.e.  97)  and 
also  by  Drummond  (313).  This  is  likewise  true  of  the  protein  of 
protein-free  milk,  as  we  have  already  seen.  It  is  necessary  to  subject 
the  protein  to  be  used  to  a  purification  process  before  it  is  fed.  In 
most  feeding  experiments  casein  is  used,  and  it  is  rendered  vitamine- 
free  by  extracting  with  alcohol  a  number  of  times,  according  to  the 
procedure  of  Funk  and  Macallum  (I.e.  81);  we  found  that  this 
procedure  did  not  decrease  the  nutritve  value  of  casein,  in  spite  of 
statements  by  McCollum  and  Davis  (I.e.  97)  to  the  contrary.  It  is, 
of  course,  possible  to  purify  the  casein  by  dissolving  it  in  alkali  and 
precipitating  with  acid,  and  also  by  continued  washing  with  water. 
In  using  meat,  according  to  Osborne,  Wakeman  and  Ferry  (I.e.  102), 
it  suffices  to  extract  it  repeatedly  with  water.  From  edestin,  as  we 
have  seen,  this  procedure  removes  the  vitamine  only  with  great 
difficulty.  Of  the  other  commonly  used  dietary  constituents,  cane 
sugar  is  vitamine-free,  as  are  also  most  of  the  varieties  of  starch. 
Regarding  fat,  we  have  used  ordinary  lard  and  under  certain  con- 
ditions it  can  be  entirely  dispensed  with.  The  purification  of  lard  is 
made  in  our  laboratory  by  autoclaving  at  30  pounds  for  3  hours ;  we 
shall  have  occasion  to  show  later  that  this  lard  is  then  vitamine-free. 
For  the  entire  dietary  mixture,  about  3  per  cent  of  agar  is  added  to 
diminish  the  possibility  of  constipation.  If  it  is  desired  to  use  a  sub- 

9  Ferry  (312)  describes  the  technique  of  rat  feeding  experiments. 
10  The  Vitamines,   1st  ed.,   p.   159. 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  113 

stance  hitherto  not  utilized,  it  is  necessary,  by  means  of  a  specific 
control,  to  test  for  the  presence  of  vitamines.  For  an  inorganic  salt 
mixture,  that  described  by  Osborne  and  Mendel,  somewhat  like  the 
inorganic  salts  of  milk,  or  the  one  used  by  McCollum  and  Simmonds 
is  available.  The  mixtures  have  the  following  composition: 

Osborne  and  Mendel  (314)  McCollum  and  Simmonds  (315} 

CaC03 134.8  NaCl 0. 173 

Na2CO3 34.2  NaH2PO4.H2O 0.347 

H3PO4 103.2  K2HPO4 0.954 

H2S04 9.2  CaH4(P04)2.H20 0. 540 

Citric  acid  •  H2O 111.  1  Iron  citrate 0. 118 

MgCO3 24.2  Calcium  lactate 1.390-1.300 

K2C03 141 . 3  MgS04  (anhydrous) 0. 266 

HC1 53.4 

Iron  citrate  •  1.5H2O  ....     6.34 

KI 0.02 

NaF 0.248 

K2A12(S04)2 0.0245 

The  food  mixture  is  best  prepared  so  that  it  has  more  or  less  of  a 
pasty  consistency,  in  which  form  it  is  least  scattered  by  the  rats. 
As  an  example  of  a  diet  now  thought  to  be  complete,  one  having  the 
following  composition  is  given: 

Substance  Per  cent        Substance  Percent 

Casein 22  Lard 20 

Cane  sugar 10  Agar 3 

Starch 27  Salt  mixture 3 

Butter 10  Yeast  extract 6 

In  older  investigations,  for  example,  in  the  work  of  Hopkins  in 
1912  (I.e.  70),  or  even  in  that  of  Funk  and  Macallum  (I.e.  86),rnot 
only  was  the  amount  of  food  consumed  taken  note  of,  but  it  was 
estimated  how  much  of  the  food  intake  was  assimilated,  by  means 
of  calorimetric  determinations  of  the  food  and  of  the  excreta.  In 
the  newer  investigations,  the  tendency  has  been  to  get  away  from  the 
above  time-consuming  procedure,  and  to  dispense  with  records  of  the 
quantity  of  food  ingested.  The  rat  has  become,  so  to  speak,  a 
reagent  for  the  testing  of  the  value  of  the  diet,  and  since  many 
thousands  of  experiments  have  been  made  in  the  last  10  years,  the 
investigators  sought  to  simplify  the  experimental  conditions  as  much 
as  possible.  Nevertheless,  the  weighing  of  the  food  seems  to  us  to 


114  THE    VITAMINES 

be  of  the  greatest  significance,  particularly  in  those  experiments 
yielding  negative  results.  It  does  not  suffice  merely  to  feed  the 
animal;  one  must  also  be  certain  that  it  eats  enough.  If  no  food 
control  is  provided  for,  it  is  easy  to  make  false  conclusions;  it  would 
be  possible  to  designate  a  certain  diet  as  insufficient,  whereas  the 
true  reason  for  the  misleading  experiments  lies  in  the  unsuitability 
of  the  animal.  This  applies  in  general  to  rats  that  are  bought;  it 
may  be  worth  while  mentioning  that  we  have  had  particularly  good 
results  with  black  and  white  rats,  raised  in  the  laboratory ;  this  strain 
seems  to  be  unusually  resistant. 

It  must  be  emphasized  once  more  that  not  all  problems  of  nutrition 
can  be  solved  by  the  use  of  rats  as  experimental  animals.  To  illus- 
trate, Osborne  and  Mendel  (316)  have  shown  that  when  the  growth 
of  rats  is  prevented  by  a  specially  chosen  diet,  growth  may  be  resumed 
after  they  have  reached  two-thirds  of  their  size,  upon  the  addition 
of  the  missing  factor.  While  Jackson  and  Stewart  (317),  in  con- 
firmation of  these  findings,  concluded  that  this  ability  to  resume 
growth,  after  a  period  of  repression,  depends  upon  the  age  of  the 
animals  and  the  duration  of  the  growth  inhibition  (in  agreement  with 
the  work  of  Briining  (318)  and  Aron  (319)  ),  and  under  certain  cir- 
cumstances may  be  appreciably  affected,  we  must  nevertheless  admit 
that  the  rat  does  possess  this  ability  for  the  most  part.  Now,  it 
would  be  futile  to  try  to  achieve  the  same  results  in  man  or  other 
mammals  of  a  corresponding  age.  As  a  matter  of  fact,  various  kinds 
of  .animals  behave  quite  differently.  In  the  same  way,  it  is  evidently 
impossible  to  study  the  nature  of  scurvy  and  pellagra  in  these 
animals  as  some  investigators  have  done.  The  individual  animals  do 
not  compare  in  regard  to  nutrition  and  nutritional  diseases. 

An  interesting  question  is  involved  in  experiments  with  inadequate 
diets  to  show  how  far  rats  have  the  power  to  choose  out  of  two  diets, 
the  one  that  is  adequate.  This  was  investigated  by  Slonacker  (320) 
with  a  primitive  method,  and  repeated  later  by  Osborne  and  Mendel 
(321).  In  the  latter  experiments,  it  could  be  shown  that  the  rats 
chose  correctly  almost  invariably. 

The  vitamine  requirements  of  rats.  Although  the  individual  phases 
of  this  question  have  been  briefly  considered  in  the  historical  part  and 
other  sections  of  the  book,  it  appears  necessary  to  treat  systematically 
this  matter  which  has  assumed  such  practical  as  well  as  theoretical 
importance.  We  have  already  said  (even  though  the  question  should 


THE    VITAMINES   IN   THE    ANIMAL   KINGDOM  115 

not  belong  to  the  subject  of  vitamines)  that  for  the  correct  feeding 
of  rats,  it  is  necessary,  above  all,  that  the  food  should  have  the  proper 
composition.  We  know  that  a  protein  is  necessary  whose  content 
of  amino  acids  is  above  reproach.  We  may  say  that  casein,  lactal- 
bumin,11  egg-albumin,  muscle  protein,  as  well  as  some  plant  proteins 
have  been  shown  to  be  adequate  for  this  type  of  animal.  Suzuki 
and  his  co-workers  (323)  demonstrated  that  an  artificial  mixture  of 
amino  acids  does  not  permit  of  growth  in  the  rat,  while  digested 
meat  (erepton)  possesses  this  ability  because  of  its  vitamine  content. 
Aside  from  this,  it  may  be  connected  with  a  more  favorable  relation- 
ship of  the  amino  acids  to  each  other.  In  addition,  we  refer  to  the 
excellent  work  of  Osborne  and  Mendel,  who  have  carefully  studied 
the  value  of  various  proteins  on  rats. 

Furthermore,  it  is  necessary  that  the  rats  receive  a  sufficient  and 
properly  composed  salt  mixture,  and  that  the  individual  constituents 
of  the  diet  are  present  in  proper  proportion.  This  point,  to  be 
mentioned  again  shortly,  is  of  great  significance.  It  cannot  be  said 
with  certainty  that  these  basic  conditions  are  always  accurately 
maintained ;  it  is  only  in  the  work  of  the  last  six  years  that  they  have 
been  given  their  true  value.  All  of  the  above  mentioned  factors  must 
be  taken  into  account  before  undertaking  a  vitamine  problem. 

We  have  already  called  attention  to  the  work  of  Osborne  and 
Mendel  in  1911-1912,  in  which  they  still  clung  to  the  idea  that  they 
had  solved  the  question  of  artificial  nutrition.  At  first  they  did  not 
see  that  the  addition  of  "protein-free  milk"  to  the  diet  was  an 
unknown  factor  whose  significance  for  nutrition  could  not  be  accu- 
rately gauged  at  the  time.  This  happened  in  spite  of  the  work  of 
Stepp,  who  had  already  affirmed  the  presence  of  essential  lipoids  in 
milk.  Through  the  classical  work  of  Hopkins  and  through  our  (324) 
simultaneous  demonstration  of  vitamine  B  in  milk,  it  became  clear 
that  something  could  adhere  to  the  products  made  from  milk;  this 
might  explain  the  striking  results  obtained  by  Osborne  and  Mendel. 
Hopkins  showed  that  a  small  quantity  of  milk  (a  few  cc.), 

11  Sure  (322)  has  recently  found  that  lactalbumin  is  not  a  complete  protein 
since  it  does  not  contain  enough  cystine  and  tyrosine.  According  to  this, 
it  is  obvious  that  the  results  of  Emmett  and  Luros  (I.e.  105),  who  suggested 
the  probability  of  a  new  vitamine,  and  who  found  that  lactalbumin  was 
rendered  complete  when  supplemented  with  protein-free-milk,  could  be 
explained  by  the  presence  of  cystine  and  tyrosine  in  the  protein-free  milk. 


116 


THE   VITAMINES 


added  to  a  carefully  purified  food  mixture,  resulted  in  normal  growth 
of  rats;  at  the  same  time  the  food  intake  increased  markedly.  The 
favorable  effect  of  these  small  amounts  of  milk  is  best  shown  in  the 
accompanying  curve.  Since  Hopkins  fed  milk  in  the  natural  state, 
its  effect  could  be  due  to  the  cumulative  influence  of  several  unknown 
factors — which  was  found  later  to  be  the  case.  Subsequently,  Funk 
(325),  and  also  Osborne  and  Mendel  (326),  showed  that  milk  does- 


FIG.  22.  GROWTH  CURVES   (HOPKINS) 

Lower  curve  (up  to  eighteenth  day),  vitamine-free  diet;  then  3  cc.  milk 
daily;  upper  curve,  the  reverse. 

not  exert  such  a  great  influence  as  Hopkins  first  found;  however,  we 
must  not  forget  that  the  vitamine  content  of  milk  and  other  food 
stuffs  does  not  represent  a  constant  mathematical  unit  and  can  under- 
go great  variations. 

McCollum  and  Davis  (I.e.  75),  and  shortly  thereafter  also  Osborne 
and  Mendel  (I.e.  76),  showed  that  p)rotein-free  milk  was  not  sufficient 
for  the  growth  of  rats,  and  that  the  diet  needed  certain  fatty  con- 
stituents, found  in  egg-yolk  and  in  butter.  The  lack  of  these  factors 


THE   VITAMINES   IN    THE   ANIMAL   KINGDOM  117 

was  evidenced  by  a  general  undernutrition,  a  rough  coat  and  a 
characteristic  eye  disease,  now  called  ophthalmia  (keratomalacia) . 
Since  this  disease  does  not  occur  only  in  rats,  we  shall  have 
occasion  to  speak  of  it  again.  It  was  difficult  to  bring  these  new 
findings  into  accord  with  the  then  prevailing  knowledge  of  vitamines, 
the  more  so  since  Osborne  and  Mendel  (I.e.  72)  obtained  good  growth 
with  fat-free  mixtures.  Later  on,  Stepp  (327)  showed  that  his 
"life-sustaining"  substances  did  not  belong  to  the  lipoids,  and  Lander 
(328)  demonstrated  that  lecithin  and  cholesterol  played  no  particular 
part  in  nutrition.  Hence  we  could  not  understand  why  the  role  of 
vitamine  B  was  relegated  to  the  background,  and  even  denied. 
Subsequently,  Aron  (329)  also  reported  on  the  importance  of  butter.12 
Although  we  had  already  at  that  time  assumed  the  existence  of  a 
special  vitamine,  associated  with  fats  (in  cod  liver  oil,  we  assumed 
the  existence  of  a  specific  antirachitic  vitamine  because  of  the  partial 
lack  of  vifcamine  B  and  the  therapeutic  influence  on  rickets)  we  were 
somewhat  skeptical  of  the  existence  of  a  vitamine  in  butter.  In  this 
we  were  strengthened  by  our  experiments  together  with  Macallum, 
particularly  since  butter  was  shown  to  be  free  from  B-vitamine  and 
since  it  was  possible  (330),  by  substituting  fresh  yeast  for  dry  yeast, 
adding  orange  juice,  and  treating  the  eyes  with  zinc  sulfate  and  boric 
acid  solution,  to  maintain  animals  in  good  condition  for  150  days. 
The  substitution  of  butter  for  lard  was  without  influence  in  our 
experiments.  All  these  results  are  now  easily  explainable,  and  yet 
we  must  assert  that  we  have  observed  instances  of  ophthalmia  on  a 
diet  containing  butter.13  Our  findings .  showed  that  the  substance 
present  in  butter  was  not  the  only  growth  factor,  asMcCollum  would 
have  it,  but  that  vitamine  B  is  at  least  quite  as  important  as  vita- 
mine  A.  This  is  well  illustrated  in  the  curve  taken  from  the  work  of 
Funk  and  Macallum. 

12  Aron  (I.e. 319)  introduced  the  method  of  giving  the  vitamines  as  a  medicine, 
so  to  speak,  independent  of  the  diet.     This  procedure  has  the  advantage 
that  the  vitamine  addition  has  nothing  to  do  with  the  food  consumption, 
although  it  is  not  always  easy  to  give  the  necessary  dose  to  the  animals,  as 
we  ourselves  can  verify. 

13  Lately,  in  association  with  Dubin  (331),  we  have  noted,  among  30  rats, 
2  that  developed   ophthalmia  on  a  diet    containing  the   usual   amount   of 
vitamine  A.     One  of  them  was  improved  by  the  addition  of  yeast  to  the  diet. 
The  other  improved  without  medication. 


118 


THE   VITAMINES 


Effect  on  rats  of  the  lack  of  vitamine  B.  If  a  diet  is  chosen,  complete 
in  everything  else  but  lacking  in  vit amines  A  and  B,  a  nitrogen 
balance  can  be  maintained  for  24  days,  according  to  the  findings  of 
Desgrez  and  Bierry  (332).  If  the  experiments  are  extended  for  a 
longer  time,  we  obtain  the  following  curve,  reproduced  in  the  Report 


20~ 


10  20  <30  fO          DAYS 

FIG.  23.  SIGNIFICANCE  OF  VITAMINE  B  FOR  GROWTH  OF  RATS 

Vitamine  A  was  present  as  butter;  addition  of  yeast  indicated  by  *  (Funk- 
Macallum). 

of  the  English  Medical  Research  Committee  (333)  on  vitamines.  It 
may  be  seen  that  growth  ceases  completely  if  both  vitamines  are 
lacking,  and  the  animals  would  have  died  if  these  factors  had  not 
been  administered  in  time.  The  addition  of  vitamine  A  only  had  no 
influence  on  the  growth,  while  the  addition  of  vitamine  B  resulted  in 
slight  growth,  which,  however,  stopped  again  after  a  short  time. 


THE    VITAMINES    IN    THE    ANIMAL   KINGDOM 


119 


The  difference  in  the  mode  of  action  of  both  vitamines  was  explained 
by  the  supposition  that  there  is  only  a  small  reserve  supply  of  the 
antiberiberi  vitamine  in  the  body,  and  hence  a  lack  of  it  is  almost 
immediately  noticeable;  on  the  other  hand,  the  reserve  of  antirachitic 
vitamine  was  thought  to  be  greater.  This  observation  may  naturally 
be  explained  in  another  way,  namely,  that  vitamine  B  is  of  greater 
significance  in  metabolic  processes  and  growth,  and  hence  larger 
amounts  are  necessary.  In  accordance  with  this,  is  the  fact  that 
grown  rats  require  less  vitamine  A  than  young  rats,  whereas  vitamine 
B  is  necessary  throughout  the  course  of  life.  When  there  is  a  lack 
of  vitamine  B,  symptoms  arise  which  have  been  likened  to  those  of 
beriberi  by  Funk  and  Macallum.  Be  that  as  it  may,  these  symptoms 


(CUW 

150 
»00 

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/^ 

s 

> 

/ 

*/ 

tf 

/ 

/ 

^ 

3* 

? 

*/ 

"'  - 

J 

^ 

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L... 

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MONTH 



++++++ 

/ 

FIG.  24.  GROWTH  OF  RATS  SHOWING  THE  DEPENDENCE  ON  VITAMINES  A  AND 
B.     (REP.  MED.  RES.  COM.) 

Diet  without  vitamine  B 

-  Complete  diet 

Diet  without  vitamines  A  and  B 

x  x  x  x  Diet  without  vitamine  B 

may  be  obviated  just  as  in  avian  beriberi.  Whether  it  really  is 
beriberi  remains  to  be  demonstrated.  A  paresis  of  the  legs  is  no 
rare  occurrence  in  these  rats.  Voegtlin  and  Lake  (334)  observed 
that  rats  are  much  more  resistant  to  beriberi  than  are  cats  and  dogs. 
Since  beriberi  is  not  an  infectious  disease,  we  cannot  speak  of  a 
resistance ;  rather  must  we  say  that  the  lack  of  vitamine  B  in  rats  is 
not  followed  by  characteristic  symptoms.  It  is  interesting  to  follow 
the  influence  of  an  addition  of  vitamine  B  upon  the  food  intake,  as  is 
shown  in  an  experiment  by  Osborne  and  Mendel  (335).  Osborneis 
of  the  opinion  that  a  daily  addition  of  0.2  gram  yeast  is  quite  suffi- 
cient to  provide  for  the  vitamine  B  requirement  of  rats.  Osborne 
and  Mendel  (336)  rightly  maintain  that  the  effect  of  vitamine  B 


120 


THE   VITAMINES 


^oo 

150 
100 

fr 

*/ 

x" 

/ 

7 

-,« 

/ 

r 

J 

/-- 

-A- 

••-»——- 

7 

^r 

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/ 

7 

MONTH 

9 

1 

O.  SHOWING  FAILURE  TO  GROW  ON  A  DIET  DEFICIENT  IN  VITAMINE 
B,  AND  RECOVERY  ON  ADDING  THAT  FACTOR  (REP.  MED.  RES.  COM.) 

Diet  without  vitamine  B 

—  Complete  diet 


160 


140 


NO 
B-VITAMIN£ 


1 2O 


NO 


100 


B-VIT 


VMI/VE 


I 


80 


FOOD 
INTAKE 


FOOD 
INTA 


40 


;to 

DAYS 


FIG.  26.  INFLUENCE  OF  VITAMINE  B  ON  THE  FOOD-INTAKE  (OSBORNE) 


THE    VITAMINES   IN   THE    ANIMAL    KINGDOM 


121 


extends  to  the  entire  metabolism,  and  that  the  specific  influence  does 
not  depend  upon  the  food  intake. 

The  effect  of  the  lack  of  vitamine  B  on  rats  was  studied  by  Drum- 
mond  (337) .  Among  other  observations,  creatinuria  was  noted ;  this 
was  connected  with  the  cleavage  of  muscle  tissue.  Grown  animals 
often  exhibited  subnormal  temperatures,  while  paralytic  symptoms 
were  rarely  noted.  The  right  heart  in  some  cases  was  enlarged  and 
the  testicles  showed  a  suppression  of  spermatogenesis.14  Emmett  and 
Allen  (339)  investigated  the  organs  and  glands  of  rats  kept  on  a  diet 
poor  in  vitamine  B.  They  noted  atrophy  of  the  thymus  and  hyper- 


£50 


.-200 


150 


100 


JO 


MONTH 


15 


7 


IVFFK 


TIG.  27.  SHOWING   PRELIMINARY  GROWTH  AND  EVENTUAL  DECLINE  ON  DIET 
DEFICIENT  IN  VITAMINE  A 

Also  recovery  on  addition  of  a  source  of  vitamine  A.  In  the  small  diagram, 
the  lower  curve  shows  growth  of  breast-fed  young  when  the  mother  is  fed  on 
a  diet  deficient  in  A-vitamine;  the  upper  curve  shows  the  same  when  the 
mother  is  fed  on  a  complete  artificial  diet.  (Rep.  Med.  Res.  Com.) 

trophy  of  the  adrenals,  similar  to  the  findings  in  birds.  Liver,  heart 
and  other  organs,  particularly  certain  intestinal  segments,  were  found 
inflamed  and  infiltrated.  Frequently,  a  fatty  degeneration  of  some 
organs  was  noted.  Lack  of  vitamine  A,  however,  led  to  no  patholog- 
ical changes,  at  any  rate  not  of  such  a  nature  as  were  easily  demon- 
strated. In  this  case,  the  symptoms  were  least  apparent  in  older 

14  Osborne  and  Mendel  (338)  held  that  yeast  as  the  sole  source  of  proteins 
causes  sterility  in  rats,  but  it  still  remains  to  corroborate  this  finding.  These 
investigators  confirmed  our  statement  (I.e. 325)  that  for  rats,  yeast  protein 
can  completely  replace  the  protein  of  the  diet. 


122  THE    VITAMINES 

animals.  The  animals  showed  a  coarse  and  sparse  coat,  and  a 
decreased  resistance  toward  infections  of  all  kinds,  such  as  ophthal- 
mia, and  pulmonary  affections,  like  pneumonia.  Tsuli  (339a) 
investigated  the  action  of  deficient  diets  on  thyroid,  gonads,  parotid, 
pancreas  and  other  organs.  Aron  (340)  observed  these  conditions, 
including  neuro-paralytic  changes  in  the  skin,  to  which  he  also 
attributed  ophthalmia.  Morgulis  and  Gies  (340a)  found  the  calcium 
content  of  rachitic  bones  and  teeth  lower  than  in  the  normal.  Mattill 
(340b)  compared  the  creatine,  creatinine,  and  urea  content  of  the 
blood  in  fasting  rats  and  those  deprived  of  B-vitamine.  The  non- 
protein  N  was  increased  in  both  cases — in  fasting  animals,  mostly  in 
the  form  of  urea,  while  in  the  others,  it  was  in  the  form  of  creatine, 
Briining  (340c)  saw  in  rats  fed  on  a  carbohydrate-rich  diet  a  patho- 
logical condition  arise  resembling  "Mehlnarhrschaden." 

A  question  that  has  greatly  interested  the  author  for  a  long  time, 
was  to  determine  if  with  both  of  the  above  mentioned  vitamines,  all 
of  the  nutritive  requirements  of  the  rat  have  been  fulfilled.  Looking 
through  the  many  reports  dealing  with  this  problem,  one  is  tempted 
to  answer  this  question  in  the  affirmative.  First  of  all,  we  must  see 
whether  the  growth  that  has  been  obtained  till  now  on  artificial 
diets,  represents  the  optimum  growth  of  rats.  It  is  obvious  from 
most  of  the  growth  curves  that  growth  on  the  experimental  diet 
compares  favorably  with  that  on  a  normal  diet.  To  satisfy  ourselves 
on  this  point,  we  (I.e.  325)  followed  the  growth  of  a  number  of  rats 
on  a  normal  diet,  which  contained,  among  other  things,  yeast  and 
condensed  milk.  The  experiments  showed  that  the  growth,  regarded 
by  Osborne  and  Mendel  and  also  McCollum  as  normal,  was  not  the 
optimum  for  this  type  of  animal.  For  comparison,  we  append  the 
normal  weight  curves  obtained  by  us,  Osborne  and  Mendel,  and 
McCollum.  It  should  also  be  noted  that  in  a  large  percentage  of 
these  investigations,  particularly  those  of  McCollum  and  his  co- 
workers,  still  more  complicated  food  complexes  were  fed,  which,  at 
least  theoretically,  might  still  contain  one  or  more  unknown  factors. 
On  the  other  hand,  we  must  admit  that  in  laboratories  greatly  experi- 
enced in  rat  nutrition,  for  example,  those  of  Osborne  and  Mendel, 
the  condition  of  the  artificially  fed  rats,  in  relation  to  the  state  of 
health  and  the  ability  to  rear  their  young,  leaves  nothing  to  be 
desired.  The  explanation  here  may  be  that  through  considerable 
experience  a  dietary  composition  has  been  chosen,  the  components 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM 


123 


of  which  by  chance  may  supplement  each  other.  In  our  earlier  work, 
we  believed  at  first  that  we  could  conclude  from  the  symptoms  (dis- 
position to  bleeding,  petechiae  and  rosary)  and  from  the  favorable 
influence  of  orange  juice,  that  we  were  dealing  with  an  antiscorbutic 
vitamine  deficiency.  Macallum  (341)  later  adopted  the  same  view. 
Recently,  there  appeared  the  papers  of  Harden  and  Zilva  (I.e.  100)  and 
Drummond  (I.e.  79)  completely  corroborating  the  above  observa- 
tions. Still,  some  doubt  has  lately  arisen,  since  Osborne  and  Mendel 


210 


180 


150 


x:\zo 
O 


90 


60 


FUNK 


OSE 
MENDEL 


)RNE 

X 


DAYS 


M'COU.UM 


FIG.  28.   COMPARISON  OF  NORMAL  RAT  GROWTH  CURVES  OF  OSBORNE  AND 
MENDEL,  McCoLLUM,  AND  FUNK 

(I.e.  92)  as  well  as  Byfield,  Daniels  and  Loughlin  (I.e.  90),  showed  that 
vitamine  B  is  found  in  appreciable  quantities  together  with  vitamine 
C  in  fruit  juices.  This  is  in  agreement  with  our  latest  observations, 
noted  in  our  work  with  Dubin,  that  rats  on  a  diet  containing  sufficient 
vitamine  nevertheless  show  improvement  upon  the  addition  of  extra, 
vitamine.  Besides,  in  our  earlier  and  later  work  we  have  often  found 
that  an  addition  of  freshly  prepared  dietary  mixture  is  followed  by 
marked  stimulation  of  growth. 


124 


THE   VITAMINES 


Reviewing  the  many  publications  dealing  with  the  growth  of  rats, 
it  is  evident  that  only  strictly  positive  or  strictly  negative  results  are 
recorded.  On  the  other  hand,  we  have  often  seen  rats  that  were 
not  in  the  best  of  health  after  90  to  100  days  on  a  diet  that  could  be 
regarded  as  complete  in  every  respect.  To  give  a  concrete  example. 


190 


170 


150 


130 


90 


NOfl 


MAL 


12..  , 
DAYS 


NUTF 


ITIOfv 


face 


MILK 


"FiG.  29.  RETARDED  GROWTH  SOMETIMES  NOTED  IN  RATS  ON  A  SUPPOSEDLY 

ADEQUATE   DIET,    COMPARED   WITH   THE   NORMAL 

CURVE    (FUNK-DUBIN). 

we  have  only  to  record  a  diet  used  in  the  work  of  Funk  and  Dubin 
(I.e.  331): 

Substance  Percent        Substance  Percent  Substance  Percent 

Extracted  meat ...  22      Lard 20      Autolyzed  yeast .  .  .  4 

Cane  sugar 15      Starch 22      Orange  juice 3 

Butter 10      Agar 3      Salts 3 

Despite  the  fact  that  this  composition  is  above  reproach,  in  the 
light  of  present-day  knowledge  of  nutrition,  our  animals  did  not 
grow  very  well.  Thereupon,  we  tried  to  bring  about  normal  growth 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  125 

by  the  addition  of  vitamines  A  and  B,  tomato  juice,  small  quantities 
of  milk,  and  by  the  substitution  of  casein  for  meat.  Although  some 
of  these  modifications  were  effective  for  a  short  time,  the  growth 
curve  was  nevertheless  flat.  That  the  animals  were  still  able  to 
grow,  was  demonstrated  by  placing  them  on  the  usual  normal  diet. 
Hopkins  (341a)  recently  also  reported  that  rats  failed  to  grow  on  ade- 
quate food  mixtures,  and  he  ascribes  this  to  the  seasonal  variation 
of  the  growth  impulse.  Osborne  and  Mendel  (I.e.  336)  also  showed 
that  it  is  not  so  easy  to  raise  rats  with  an  addition  of  yeast  as  with 
the  addition  of  protein-free  milk;  it  was  particularly  noticeable  that 
there  was  a  difference  in  the  ability  of  the  rats  to  raise  their  young. 
Sugiura  and  Benedict  (I.e.  108)  obtained  similar  results,  as  did  also 
Freise  (342),  who  ascribed  a  particular  role  to  milk. 

In  collaboration  with  Dubin  we  (342a)  have  recently  demonstrated 
that  what  has  up  to  now  been  known  as  vitamine  B  does  not  appear 
to  be  sufficient  for  the  growth  of  rats.  Thus,  it  seems  that  autolyzed 
yeast  contains  besides  vitamine  B  something  else  of  a  vitamine  nature 
without  which  the  rat  does  not  grow.  This  will  be  discussed  more  in 
detail  in  the  chapter  on  the  " Chemistry  of  Vitamine  B."  Judging 
from  the  data  recorded  here,  it  appears  desirable  to  study  the  nutri- 
tive requirements  of  rats  still  further. 

Mice 

As  we  have  already  seen,  the  pioneer  work  of  Stepp,  Rohmann, 
Lunin,  Socin  and  others  on  artificial  feeding  was  done  with  mice. 
Getting  away  from  the  older  work,  which  has  more  of  a  historical 
interest,  and  coming  to  the  modern  investigations,  we  see  in  the  work 
of  Brailsford  Robertson  (343),  statistical  data  in  relation  to  growth 
of  mice,  which  could  be  useful  in  judging  the  effect  of  experimental 
diets.  Thompson  and  Mendel  (344)  studied  the  food  requirements 
necessary  for  the  growth  and  maintenance  of  the  mouse,  the  suppres- 
sion of  growth,  shown  by  undernourishment,  and  the  amount  of  food 
necessary  to  overcome  this  suppression.  Mendel  and  Judson  (345) 
working  with  mice  analyzed  the  total  dry  substance,  fat  and  ash  of 
various  dietary  constituents.  M.  B.  Schmidt  (346)  made  the  inter- 
esting observation  that  if  a  deficient  diet  has  no  effect  on  the  first 
generation  in  these  animals,  its  results  are  apparent  in  subsequent 
generations.  Morpurgo  and  Satta  (347)  worked  with  mice  in 
parabiosis  in  which  one  animal  received  only  cane  sugar  while  the 


126  THE    VITAMINES 

other  was  given  a  complete  diet.  Both  mice  were  kept  alive,  showing 
that  the  one  animal  received  with  blood,  not  only  the  usual  dietary 
constituents,  but  also  the  vitamines. 

Of  the  important  investigations  on  mice,  we  shall  first  mention 
those  of  Stepp,  upon  whose  work  of  1909-1911  we  have  already 
touched  lightly.  He  extracted  the  whole  diet,  thereby  partly  remov- 
ing and  partly  destroying  the  vitamines.  In  this  way,  he  obtained 
mixed  avitaminoses  that  could  be  attributed  to  the  lack  of  at  least 
two  vitamines.  He  proved  (348)  that  lecithin,  cholesterol,  kephalin 
cerebron  and  phytin  could  not  replace  the  substances  extracted  by 
alcohol-ether  mixture.  He  found  also  that  while  the  important  sub- 
stances could  not  be  removed  by  means  of  the  ether  extraction  alone ; 
they  could  be  removed  with  alcohol  alone.  When  egg-yolk  is 
extracted  with  alcohol,  these  substances  go  into  solution,  but  not 
when  acetone  is  used.  Following  this,  Rohl  (349)  found  that  ordinary 
commercial  lecithin  contained  these  substances,  while  pure  lecithin 
did  not.  In  a  publication  appearing  in  1914,  Stepp  (350)  drew  close 
to  the  prevailing  view  of  the  importance  of  two  vitamines  in  the  life 
of  mice  although  he  still  thought  of  the  fat  vitamine  as  a  lipoid. 
In  1916,  Stepp  (351)  advanced  a  step  further  in  that  he  used  two 
different  extracts,  one  of  egg-yolk  and  the  other  of  rice  polishings 
(orypan).  However,  the  experiments  in  this  case  were  not  very 
clear,  since  ab  first  Stepp  worked  with  a  vitamine  mixture  (egg-yolk), 
believing  he  had  to  deal  with  a  lipoid.  Furthermore,  he  showed  (352) 
that  a  mixture  of  lecithin,  kephalin,  cerebron,  cholesterol  and  orypan, 
added  to  dog  biscuit  which  had  been  extracted  with  alcohol,  could 
at  least  maintain  mice  at  a  certain  level.  It  is,  however,  not  impos- 
sible that  this  was  due  to  a  cumulative  action  of  vitamine  A  con- 
tamination. Dezani  (353)  conducted  experiments  for  the  purpose 
of  determining  the  source  of  cholesterol.  He  fed  mice  on  flour  and 
casein,  extracted  with  alcohol  and  ether.  The  animals  died  after 
18  to  19  days,  showing  a  41  per  cent  loss  in  weight.  Cholesterol 
was  found  in  the  feces,  and  Dezani  (354)  was  of  the  opinion  that  mice 
could  be  kept  alive  on  a  lipoid-free  diet.  We  may  mention  here  one 
of  the  typical  experiments  of  Rohmann  (355),  in  which  mice  were 
raised  on  a  diet  of  casein,  chicken  protein,  nucleoprotein  from  liver, 
potato  starch,  margarine  and  a  salt  mixture.  This  food  proved  to 
be  inadequate  and  the  animals  grew  less  than  on  milk  and  rolls.  The 
substitution  of  vitellin  for  nucleoprotein  did  not  yield  any  better 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM  127 

results,  and  Rohmann  regarded  this  diet  as  adequate  only  for  grown 
animals.  Meat  extract  and  dried  meat  powder  had  no  particularly 
favorable  effect;  egg-yolk  on  the  contrary,  was  favorable  but  not 
sufficient  for  growth,  while  egg-fat  was  not  well  tolerated,  even  in 
older  animals.  In  a  second  series  of  experiments,  the  food  was 
supplemented  with  yeast  and  baked,  with  the  addition  of  egg-yolk. 
The  results  were  very  good  and  were  attributed  to  the  yeast ;  lecithin 
was  shown  to  be  unnecessary.  It  may  be  seen  that  Rohmann's 
findings  coincide  with  those  on  rats.  Wheeler  (356)  carried  out  some 
experiments  with  mice,  planned  in  accord  with  the  first  experiments 
of  Osborne  and  Mendel  on  rats.  Here,  too,  protein-free  milk  was 
used,  and  found  insufficient  as  the  sole  source  of  vitamine;  artificial 
protein-free  milk  proved  to  be  entirely  inadequate.  It  seems  that 
mice  need  more  protein  than  do  rats  in  order  to  grow;  yet  the  former 
grow  more  rapidly  after  the  addition  of  a  milk  preparation  than  do 
the  latter.  MacArthur  and  Luckett  (357)  investigated  the  influence 
of  lecithin,  kephalin,  cerebrosides  and  cholesterol  as  additions  to  an 
artificially  prepared  diet,  and  came  to  the  conclusion  that  these 
substances  are  superfluous.  Furthermore,  they  studied  the  influence 
of  egg-yolk  and  found  that  the  necessary  substances  (apparently 
vitamines,  as  they  remarked  at  the  time)  are  insoluble  in  ether, 
soluble  in  cold  alcohol  and  the rmo-st able.  Butter  and  olive  oil  were 
inactive,  without  any  other  additions.  Mitchell  and  Nelson  (358) 
found  that  mice  do  not  grow  on  an  artificial  diet  till  protein-free  milk 
was  added.  From  the  foregoing,  it  is  evident  that  mice  behave  like 
rats  in  their  vitamine  requirements. 

Guinea  pigs 

Next  to  the  pigeon  and  the  rat,  the  guinea  pig  is  one  of  the  most 
widely  used  animals  for  the  purpose  of  vitamine  research,  especially' 
in  the  investigation  of  scurvy.  It  has  been  shown  unsuited  for  other 
avitaminoses,  though  there  are  no  lack  of  investigators  who  propose 
the  use  of  the  guinea  pig  for  the  study  of  pellagra  and  rickets,  despite 
its  demonstrated  unsuit ability.  This  animal  is  likewise  little  used 
for  beriberi  research  although  Schaumann  (I.e.  2)  reported,  out  of 
65  guinea  pigs  fed  on  dry  grain,  two  typical  cases  of  beriberi,  with 
subsequent  histological  demonstrations  of  multiple  neuritis.  These 
results  have  never  been  confirmed.  On  the  whole,  it  is  not  easy 
to  conduct  planned  feeding  experiments  with  these  animals,  and 


128  THE   VITAMINES 

hence  the  difficulty  of  studying  problems  of  nutrition.  We  know 
comparatively  little  of  their  vitamine  requirements,  particularly  as 
to  vitamines  A  and  B,  since  in  the  study  of  experimental  scurvy, 
dietary  mixtures  are  used  containing  both  of  these  vitamines.  Hume 
(358a)  has  recently  found  that  guinea  pigs  require  vitamine  A  and 
hence  these  animals  can  be  used  for  the  detection  of  vitamine  A  in 
green  foods  but  not  in  fats.  Tozer  (358b) ,  in  a  pathological  investi- 
gation of  these  animals  could  not  find  any  difference  between  experi- 
mental scurvy  and  the  changes  produced  by  lack  of  vitamine  A. 


FIG.  30.  LONGITUDINAL  SECTION   or  A  RIB  OF  A  SCORBUTIC  GUINEA  PIG,. 
WITH  FISSURES,  HEMORRHAGES  IN  THE  PERIOSTEUM  AND  PALE 
MARROW  (HOLST-FROLICH) 

We  owe  the  discovery  of  experimental  scurvy  to  Hoist  and  Frolich 
(359)  in  1907,  although  in  1895  a  similar  condition  was  observed  in 
the  same  species  by  Theobald  Smith  (360).  Hoist  and  Frolich 
investigated  scurvy  in  various  animals  and  chose  the  guinea  pig  as 
most  suitable  for  this  purpose.  It  appeared  to  be  unnecessaiy  to 
choose  a  special  diet,  for  these  investigators  noted  that  scurvy  could 
be  produced  in  guinea  pigs  with  any  diet  that  would  do  the  same 
in  man;  this  was  also  the  case  with  any  diet  that  was  autoclavedat 
a  high  temperature.  They  recognized,  with  great  perspicacity,  that 
the  symptoms  were  closely  analogous  to  those  in  man.  Guinea  pigs 


THE    VITAMINES    IN    THE    ANIMAL    KINGDOM 


129 


fed  with  rye,  or  wheat  bread  (and  water),  or  with  oats,  rye,  wheat, 
barley,  and  rice  flour,  died  within  a  few  weeks.  Hoist  and  Frolich 
used  animals  weighing  about  350  grams  since  they  could  better  with- 
stand the  great  loss  in  weight  (about  40  per  cent) .  In  a  later  publica- 
tion, the  same  investigators  (359)  came  to  the  conclusion  that  human, 
as  well  as  guinea  pig,  scurvy  may  be  attributed  to  the  same  causes; 
they  sought  also  to  differentiate  between  experimental  scurvy  and 
simple  inanition.  Starvation,  in  which  there  is  a  loss  of  from  30  to 
40  per  cent  in  weight,  yields  no  scorbutic  symptoms;  on  the  contrary, 
scorbutic  symptoms  ordinarily  appear  before  the 
animals  show  any  appreciable  emaciation. 

Not  all  workers  accept  the  simple  etiological  ex- 
planation of  scurvy  in  guinea  pigs.  Jackson  and 
Moore  (361)  and  Jackson  and  Moody  (362)  be- 
lieved that  there  was  an  infectious  factor  in  the 
etiology.  Moore  and  Jackson  (363)  strengthened 
their  belief  after  they  had  seen  that  raw  milk 
could  not  protect  these  animals  from  scurvy. 
The  explanation  for  this  is  to  be  sought  in  the 
probability  that  the  milk  used  did  not  contain  FIG.  31.  SCORBUTIC  FE- 
enough  vitamine  C  for  this  type  animal.  In  con-  MUR  OF  A  GUINEA  PIG, 
trast  to  the  above  investigators,  Givens  and  Hoff- 
man (364)  were  able  to  demonstrate  the  absolute 
sterility  of  the  blood  of  the  diseased  animals. 
Further,  McCollum  and  Pitz  (I.e.  69)  believed  that 
scurvy  in  guinea  pigs  was  due  to  constipation,  and  made  the 
sweeping  statement  that  scurvy,  as  an  avitaminosis,  was  non-existent, 
and  that  the  known  antiscorbutic  properties  of  fruit  juices  could  be 
explained  by  their  laxative  action.  They  stated  that  other  laxatives, 
like  phenolphthalein  and  mineral  oil,  could  be  used  with  the  same 
success.  These  statements  were  further  developed  by  Pitz  (365)  a 
co-worker  of  McCollum's,  who  found  that  lactose  acted  favorably, 
and  that  its  function  was  to  modify  the  bacterial  intestinal  flora. 
The  observations  of  McCollum  and  his  co-workers  immediately 
aroused  a  storm  of  protest.15  Shortly  thereafter,  Harden  and  Zilva 
(367)  showed  beyond  a  doubt  that  the  results  of  McCollum  were 
due  to  the  fact  that  he  gave  the  guinea  pigs  milk,  ad  libitum,  and 

15  Torrey  and  Hess  (366)  have  investigated  the  bacterial  intestinal  flora 
in  guinea  pigs  and  in  infants,  but  failed  to  find  any  abnormalities. 


WITH  PALE  MARROW 
AND  ATROPHIC 

TRABECULAE 
(HOLST-FROLICH) 


130  THE   VITAMINES 

in  this  manner  prevented  the  onset  of  scurvy.  When  milk  was 
•omitted,  lactose  had  not  the  least  influence  upon  the  progress  of 
the  disease.  Later,  these  observations  were  confirmed  by  many 
others,  and  McCollum  himself  abandoned  his  view. 

When  we  (368)  spoke  with  doubt  as  to  the  identity  of  scurvy  in 
guinea  pigs  and  in  man,  it  was  based  upon  failure  to  obtain  ther- 
apeutic results  with  milk,  lime  juice,  potato  and  lemon  juice.  All  of 
the  difficulties  mentioned  by  us  at  the  time  have  been  overcome  by 
countless  investigators,  and  at  present  there  is  no  reason  to  doubt 
that  in  guinea  pigs,  we  are  actually  dealing  with  scurvy. 

As  we  have  already  said,  the  vitamine  requirements  of  the  guinea 
pig,  aside  from  vitamine  C,  have  been  insufficiently  investigated. 
Hausermann  (I.e.  13)  extended  his  experiments  on  the  influence  of 
iron  salts  on  guinea  pigs,  without  noting  any  results.  Heim  (369) 
found  that  guinea  pigs  could  not  live  on  raw  or  cooked  cow's  milk. 
Of  eight  animals,  one  died  after  a  month,  two  after  2  months,  and  the 
other  five  remained  alive  for  3  months  (the  whole  experimental 
period),  but  showed  loss  of  weight.  In  another  series  of  experiments, 
the  animals  died  in  from  4  to  14  days;  the  addition  of  cellulose  was 
without  effect,while  malt  extract  and  alcoholic  extract  of  germinated 
barley  exerted  a  definite  favorable  influence.  Rondoni  andMontag- 
nani  (370)  conducted  similar  experiments  with  corn  and' oats,  and 
with  complete  starvation.  Guinea  pigs  on  corn  showed  milder 
scurvy  symptoms  than  those  on  oats.  Of  particular  interest,  is  the 
work  of  Ingier  (371)  who  investigated  the  influence  of  diet  on 
pregnant  guinea  pigs.  Here,  too,  a  diet  of  oats  and  water  showed 
scorbutic  influence  on  the  embryo.  When  this  diet  was  used  in  the 
later  stages  of  pregnancy,  no  pathological  changes  were  found  in  the 
bones.  A  short  feeding  with  the  milk  of  the  scorbutic  mother,  con- 
verted the  latent  scurvy  into  the  active  form.  In  pregnant  animals, 
the  disease  developed  more  rapidly,  with  more  marked  symptoms, 
than  in  those  that  were  not  pregnant,  and  in  the  first  stages  of  preg- 
nancy lead  to  death.  Ingier  described  a  fatty  degeneration  of  the 
nerves  in  these  guinea  pigs. 

Conditions  for  the  demonstration  of  experimental  scurvy. — The 
foodstuff  mostly  used  for  the  development  of  this  disease  is  oats. 
According  to  the  investigations  of  McCollum,  Simmonds  and  Pitz 
(372),  oats  are  poor  in  adequate  protein,  salts  and  vitamine  A.  It  is 
undoubtedly  true  that  under  the  working  conditions  maintained  by 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  131 

Hoist,  the  guinea  pigs  soon  lost  their  appetite  and  died,  presenting 
mixed  symptoms  of  scurvy  and  starvation.  Therefore,  it  signified  a 
step  forward  when,  according  to  the  procedure  of  Chick  and  Hume 
(373),  the  animals  were  given  an  addition  of  sterilized  milk.  This 
addition  did  not  delay  the  development  of  the  disease — the  general 
condition  of  the  animals  being  markedly  improved — and  corre- 
sponded with  the  view  advanced  by  McCollum  and  his  school  as  to 
the  nutritive  value  of  oats.  The  demonstration  of  experimental 
scurvy  was  no  longer  difficult,  especially  after  the  introduction  of  the 
milk  addition,  and  investigations  on  this  subject  were  easily  carried 
out.  For  this  purpose,  Cohen  and  Mendel  (374)  used  animals 
weighing  from  110  to  250  grams.  They  believed  that  cod  liver  oil 
had  no  influence  on  scurvy,  so  that  a  complication  of  rickets  was 
no  cause  for  concern.  Scurvy  can  be  produced  by  feeding  soya  bean 
flour,  which  contains  enough  of  vitamines  A  and  B,  supplemented 
with  cellulose  and  salts.  Hess  and  linger  (375)  used  oats,  hay  and 
water  (to  this  may  also  be  added  cod  liver  oil)  as  a  basal  dietand 
found  that  guinea  pigs  weighing  from  200  to  300  grams  developed 
scurvy  in  two  to  three  weeks. 

In  the  Lister  Institute  in  London,  where  many  investigations  on 
guinea  pigs  are  being  made,  the  original  method  of  Hoist  and  Frolich 
was  modified  so  that  the  diet  consisted  of  oats,  bran  and  60  cc.  of 
milk  for  guinea  pigs  weighing  350  grams.  The  milk  is  autoclaved 
for  one  hour  at  120°C.;  on  this  diet  the  animals  grow  normally  for 
about  15  to  20  days,  after  which  the  first  symptoms  of  scurvy  are 
noticed.  The  animals  lose  weight,  and  after  30  to  40  days  they  die 
of  acute  scurvy.  In  the  presence  of  sufficient  vitamine  C,  growth  is 
uninterrupted. 

The  symptoms,  already  largely  described  by  Hoist  and  Frolich, 
are  as  follows :  the  molars  become  loose,  with  bleeding  of  the  gums. 
Sometimes  a  bluish  hyperemia  is  noticed  and  in  rare  cases,  ulceration. 
Apart  from  this,  hemorrhages  are  common  in  the  soft  parts  of  the 
knee  joints  and  under  the  periosteum  of  the  sternal  ends  of  the  ribs. 
Very  often  the  connection  between  the  ribs  and  the  cartilage  is 
loosened;  there  is  a  severing  between  the  epiphysis  and  the  shaft  of 
the  bones,  especially  the  tibia.  Later  there  occur  duodenal  ulcers, 
brittleness  of  the  bones,  hematuria  and  edema.  In  addition  to  these 
symptoms  Chick  and  her  co-workers  (376)  observed  extreme  pain  and 
swellings  of  the  joints,  even  in  the  first  stages.  The  animal  assumes 


132 


THE   VITAMINES 


a  characteristic  posture  ("scurvy  position"),  lying  on  one  side,  the 
painful  limb  stretched  out  into  the  air.  In  other  cases,  the  animal 
lies  with  its  head  on  the  floor  of  the  cage,  a  position  caused  by  the 
pain  in  the  jaws  and  gums  ("scurvy  face-ache  position").  As  soon 
as  the  teeth  become  loose,  the  animals  refuse  their  food  and  death 
usually  follows  within  the  next  few  days.  If  the  animals  receive  not 
quite  enough  vitamine  C,  they  recover  and  begin  to  grow,  but  the 
joints  do  not  become  normal.  An  abundant  addition  of  vitamine 
C  before  the  beginning  of  the  experiment  has  no  effect  on  the 
time  of  development  of  scurvy,  according  to  Hess  (377);  hence  it 
would  seem  that  the  body  has  no  reserve  supply  of  this  vitamine. 

The  post-mortem  re- 
veals rarification  of  the 
long  bones,  with  fractures 
between  the  epiphyses  and 
the  bones  themselves;  frac- 
tures are  also  evident  in 
the  ribs.  The  bones  show 
arrested  ossification,  and 
atrophy  of  the  existing 
bone  tissue  and  of  the 
ossification  areas ;  the  bone 
marrow  loses  its  lymphoid 
character  at  the  diaphyseal 
ends,  becomes  poor  in  cells, 
and  sometimes  presents  a 
homogeneous  appearance  ("Helles  Mark").  Hess  and  linger  (378) 
called  attention  to  their  observation  of  rosary  in  scurvy,  which 
might  be  mistaken  for  rickets.  Rondoni  and  Montagnani  (I.e. 370) 
described  pathological  investigations  of  guinea  pigs  fed  on  corn. 
The  symptoms  seemed  to  resemble  those  of  experimental  scurvy 
although  they  were  more  mild.  The  organs  mostly  found  altered 
were  the  spleen,  thyroid  gland  and  adrenals;  the  central  nervous 
system,  liver,  digestive  tract,  kidney  and  bone  marrow  were  slightly 
changed,  while  the  myocardium  and  the  lungs  were  still  less 
affected.  The  changes  in  the  thyroid  and  adrenals  go  hand 
in  hand  with  a  hypertrophy  of  the  islands  of  Langerhans.  Thy- 
roid and  spleen  gave  evidence  of  sclerosis;  the  adrenals,  a  diminu- 
tion of  lipoids;  and  in  the  cortex,  atrophy  and  degeneration. 


FIG.  32.  "FACE-ACHE"  POSITION  OF  SCORBUTIC 
GUINEA  PIG  (REP.  MED.  RES.  COM.) 


THE    VITAMINES    IN   THE    ANIMAL    KINGDOM  133 

Rondoni  (379)  had  already  observed  that  the  adrenals  were  somewhat 
enlarged,  though  containing  less  adrenaline.  This  signifies  a  great 
difference  between  scurvy  and  beriberi,  for  in  the  latter,  the  amount 
of  adrenaline  is  increased.  Rondoni  believed  that  the  picture  of 
"maidismus"  (corn-feeding)  is  different  in  scurvy  and  pellagra. 
McCarrison  (380)  studied  the  influence  of  a  diet  of  oats  and  auto- 
claved  milk.  While  the  weight  of  the  normal  adrenals  in  guinea  pigs 
is  about  0.467  gram,  that  in  scurvy  was  about  0.955  gram.  Histo- 
logical  examination  revealed  hemorrhagic  infiltration  (which  perhaps 
causes  an  enlargement  of  the  gland)  and  atrophy  of  the  cellular 
elements  of  the  cortex  and  medulla.  In  spite  of  the  size  of  the 
gland,  the  amount  of  adrenaline  is  diminished  and  has  about  half  of 
its  normal  value.  The  quantity  of  adrenaline  per  gram  of  body 
weight  dropped  to  about  one-quarter  of  the  normal.  These  symp- 
toms arise  before  the  other  symptoms  are  apparent.  LaMer  and 
Campbell  (381),  working  in  Sherman's  laboratory,  were  able  to  con- 
firm the  results  of  McCarrison  completely.  The  thyroid  gland, 
according  to  McCarrison  (I.e.  295),  may  also  be  found  increased  in 
size,  due  to  infiltration.  Aside  from  this,  McCarrison  (382)  found 
that  the  bladder  had  undergone  pathological  changes.  Inflammation 
of  the  mucosa  and  the  musculosa  of  the  bladder  and  signs  of  degenera- 
tion of  the  epithelium  were  noted,  associated  with  hemorrhagic 
infiltration. 

As  regards  the  metabolism  in  scurvy,  we  have  the  data  of  Bauman 
and  Howard  (383),  differentiating  it  from  simple  starvation.  The 
animal  were  fed  on  oats,  and  the  inorganic  metabolism  was 
controlled.  Nitrogen,  sulphur  and  phosphorus  elimination  were 
decreased,  sodium  and  chlorine  were  unaffected,  potassium  was 
retained,  while  the  calcium  and  magnesium  output  was  increased. 
These  investigators  observed  the  typical  " white  line"  of  infantile 
scurvy,  noted  by  Frankel.  In  comparison,  there  are  the  normal 
figures  obtained  by  Smith  and  Lewis  (384)  with  guinea  pigs. 

Through  a  determination  of  the  alkali  reserve  of  the  blood  of  these 
animals,  McClendon,  Cole,  Engstrand  and  Middlekauff  (385)  showed 
that  scurvy  of  guinea  pigs  has  nothing  in  common  with  acidosis. 
Funk  came  to  the  same  conclusion  somewhat  earlier  (I.e.  368),  on 
the  addition  of  sodium  bicarbonate. 

Lewis  and  Karr  (386)  found  the  blood  and  organs  of  scorbutic 
animals  much  richer  in  urea,  as  compared  with  those  of  the  controls; 


134  THE   VITAMINES 

this  could  not  be  explained  by  hunger  or  loss  of  water.  Karr  and 
Lewis  (387)  studied  the  amount  of  conjugated  phenols  excreted.  The 
figures  obtained  were  normal  and  hence  their  conclusion,  that  in 
scorbutic  guinea  pigs  there  is  an  abnormal  bacterial  decomposition 
in  the  intestine,  seems  unjustified.  According  to  MeCollum  and 
Parsons  (387a),  the  prairie  dog  behaves  not  like  the  guinea  pig  but 
like  the  rat,  in  its  vitamine  C  requirements. 

Rabbits 

Rabbits  have  been  infrequently  used  in  the  solution  of  some  of  the 
vitamine  problems.  The  reason  for  this  is  that  these  animals  are 
much  less  affected  by  a  lack  of  vitamines,  which  we  have  personally 
demonstrated  (I.e.  368).  We  found  that  a  rabbit  may  live  on  oats 
alone  for  more  than  three  months,  exhibiting  individual  variations  in 
their  behavior.  The  explanation  for  this  may  be  found  in  a  paper 
by  Portier  and  Randoin  (I.e.  217).  In  this  investigation,  rabbits 
were  fed  on  cabbage  and  carrots,  autoclaved  for  one  hour  at  125°C. 
The  animals  usually  died  after  11  to  17  days,  with  symptoms  of  an 
avitaminosis.  When  the  experiments  were  repeated  with  a  larger 
animal,  these  results  could  no  longer  be  obtained;  the  animal  was 
observed  for  three  months  during  which  time  it  gained  weight.  This 
observation  found  no  explanation,  till  it  was  noted  one  day,  that  the 
animal  was  eating  the  feces.  Two  series  of  experiments  were  then 
arranged,  in  one  of  which  the  animals  received,  in  addition  to  the 
diet,  the  feces  of  the  other,  kept  also  on  the  de-vitaminized  food.  The 
animals  receiving  feces  were  in  good  health  after  100  days  and  gained 
weight.  The  investigators  explained  this  result  by  saying  that  with 
the  feces,  bacteria  were  ingested  which  prepared  vitamine  for  the 
animal  by  symbiosis.  It  must  nevertheless  not  be  forgotten,  that 
feces  may  contain  vitamine,  as  we  shall  see  later  on.  Nevertheless, 
the  observation  of  Portier  and  Randoin  was  of  greatest  interest. 

Schaumann  (I.e.  2)  thought  that  he  had  produced  beriberi  in 
rabbits,  fed  on  corn,  which  could  be  cured  by  yeast  or  Katjang-idjoe 
beans.  Since  corn  is  very  rich  in  vitamine  B,  this  condition  was 
evidently  confused  with  scurvy.  Abderhalden  and  Lampe  (I.e.  25) 
have  observed  paresis  in  a  rabbit  fed  with  rice,  but  beriberi  has  never 
been  actually  demonstrated  in  rabbits.  On  the  other  hand,  scurvy 
in  rabbits  has  been  described  by  Hoist  and  Frolich  (I.e.  36),  identical 
with  scurvy  in  guinea  pigs.  In  opposition  to  this,  Morgen  and 


THE    VITAMINES    IN   THE    ANIMAL    KINGDOM  135 

Beger  (388)  regarded  the  disease  of  rabbits  fed  on  oats  as  an  acidosis 
which  could  be  obviated  by  the  addition  of  calcium  carbonate  and 
sodium  bicarbonate.  With  these  additions,  some  of  the  animals 
lived  up  to  190  days,  and  often  showed  a  gain  in  weight.  We  (I.e. 
368)  have  repeated  this  work  and  have  satisfied  ourselves  that  the 
addition  of  alkali  actually  does  excite  the  appetite  and  prolong  life, 
but  does  not  protect  from  death.  McClendon,  Cole,  Engstrand  and 
Middlekauff  (I.e.  385)  obtained  the  same  results.  Kurijama  (389) 
found  that  the  alkali  reserve  of  the  blood  depends  very  much  upon 
whether  the  food  gives  rise  to  bases  or  acids,  although  McClendon, 
v.  Meysenbug,  Engstrand  and  King  (390)  rightly  observed  that 
his  results  were  complicated  by  scurvy,  since  he  fed  his  rabbits  on 
oats. 

From  what  has  just  been  said,  we  see  that  rabbits  are  hardly  the 
proper  animals  to  use  for  the  study  of  scurvy.  According  to  the  work 
of  Nelson  and  Lamb  (391)  rabbits  seem  to  be  adapted  for  the  study 
of  ophthalmia.  Two  rabbits,  kept  on  commercial  casein,  dextrin, 
lactose,  wheat  germ,  salts  and  an  alcoholic  extract  of  alfalfa  grass, 
developed,  after  60  days,  an  eye  disease  which  they  thought  to  be 
similar  to  ophthalmia;  one  of  the  animals  was  cured  on  the  addi- 
tion of  butter. 

Cats 

Schaumann  (I.e.  2)  fed  two  cats  on  denatured  meat  heated  in  an 
autoclave  with  sodium  carbonate  at  120°C.  One  of  the  animals 
died  after  42  days,  the  other,  after  58  days  with  complete  paralysis. 
It  seems,  according  to  the  newer  investigations,  that  what  Schaumann 
saw  was  really  beriberi.  The  experiments  were  repeated  by  Weill, 
Mouriquand  and  Michel  (392)  and  they  showed  that  the  same 
symptoms  could  be  obtained  with  cooked  as  well  as  with  sterilized 
meat.  While  the  animals  died  after  45  days  on  small  amounts  of 
fresh  meat,  nervous  symptoms  appeared  after  35  to  39  days  with 
sterilized  meat.  Voegtlin  and  Lake  (393)  produced  typical  beriberi 
in  cats  on  fat-free  meat,  digested  with  10  per  cent  sodium  carbonate 
solution  for  three  hours  at  120°C.  The  beriberi  symptoms  were 
completely  removed  with  vitamine  B,  sometimes  after  only  12  hours. 
The  disease  could  be  prevented  by  daily  additions  of  2  cc.  of  autolyzed 
yeast  per  kilo  body  weight,  but  not  by  the  addition  of  5  per  cent 
butter  or  10  per  cent  purified  casein.  On  beef,  heated  without  soda, 


136  THE   VITAMINES 

the  animals  remained  healthy,  except  during  pregnancy  and  lactation, 
when  the  vitamine  requirements  are  greater.  Osborne,  Wakeman  and 
Ferry  (I.e.  102)  found  very  little  vitamine  B  in  lean  beef,  and  noted 
that  cooking  several  times  in  water  removes  this  substance,  leaving 
only  traces.  Mackay  (393a)  and  Tozer  (393b)  did  not  succeed  in 
producing  experimental  rickets  in  cats  in  the  absence  of  vitamine  A. 

Lions 

Bland-Sutton  (394)  observed  rickets  in  young  lions  in  captivity 
when  they  are  weaned  too  soon  and  fed  on  raw  meat.  Apparently, 
there  is  a  lack  of  vitamine  A,  since  the  animals  were  cured  with  milk 
and  cod  liver  oil. 

Dogs 

Dogs  are  now  frequently  used  for  vitamine  studies  and  therefore 
we  shall  devote  a  little  more  space  to  this  subject.  The  dog  has 
become  particularly  important  for  experiments  on  rickets.  In  the 
historical  section,  we  have  already  discussed  the  metabolism  of  dogs 
to  some  extent.  It  frequently  happened,  in  these  experiments,  that 
the  various  investigators  used  a  diet  poor  in  vitamine  although  the 
purpose  of  the  experiments  was  aside  from  the  question  under  con- 
sideration. For  instance,  there  is  the  work  of  Cahn  (395)  in  1886, 
who  studied  the  results  of  chlorine  hunger.  Cooked  meat  was 
used  as  a  food,  and  Cahn  described  a  number  of  symptoms  of  chlorine 
hunger,  among  which  was  blood  in  the  stomach  contents,  which  might 
perhaps  have  been  due  to  decreased  capillary  resistance.  Similar 
experiments  were  also  carried  out  by  Trappe  (396)  with  extracted 
meat,  to  study  the  significance  of  hydrochloric  acid  in  the  stomach. 
Rosemann  (397)  too,  experimented  with  the  same  end  in  view,  as  did 
also  Bonniger  (398),  using  a  diet  of  extracted  meat.  Bonniger 
observed  that  .the  animals  refused  the  food  and  manifested  vomiting, 
apathy  and  distress.  It  was  indeed  remarkable  that  these  symptoms 
should  have  disappeared  on  the  addition  of  sodium  bromide.  Pfluger 
(399)  observed  enteritis  in  dogs  fed  on  sterilized  meat,  poor  in  fat. 

A  whole  series  of  experiments,  which  have  only  historical  interest 
at  present,  were  made  by  Wolfgang  Heubner  (400)  and  his  school 
(Lipschiitz,  Durlach)  on  the  significance  of  phosphorus  in  dogs. 
The  diet  usually  used  for  this  purpose  consisted  of  tapioca  or  white 
rice,  egg-albumin,  palmin,  cane  sugar  and  salts.  Heubner  thought 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  137 

to  obtain  a  phosphorus-free  diet  in  this  way,  whereas  in  reality,  he 
was  dealing  with  a  vitamine-free  diet.  Influenced  by  our  work, 
Heubner  later  recognized  the  importance  of  the  vitamines.  From  the 
work  of  Schmorl  (401)  who  undertook  the  pathological  examination 
of  these  dogs,  it  may  perhaps  be  noted  that  Heubner  was  dealing 
with  a  mixed  avitaminosis.  In  young  dogs,  rachitic  symptoms  pre- 
vailed, although  it  is  not  clear  whether  the  bone  changes  were  of 
scorbutic  or  rachitic  origin,  or  due  to  a  combination  of  both. 

Still  less  satisfactory  were  the  investigations  of  Masslow  (402), 
who  studied  the  same  problem  as  Heubner,  but  on  dogs,  and  on 
practically  the  same  diet.  Although  the  work  was  done  in  1913, 
Masslow  evidently  knew  nothing  of  the  significance  of  the  vitamines. 
His  only  observation  of  interest  to  us  was  the  general  emaciation  of 
the  animal  body  on  a  diet  actually  free  of  vitamines. 

In  discussing  vitamine  problems,  we  must  note,  first  of  all,  that 
dogs  seem  to  be  sensitive  to  a  lack  of  all  three  vitamine  types.  Never- 
theless, although  Hoist  and  Frolich  (I.e.  36)  produced  experimental 
scurvy  in  dogs,  they  seemed  to  be  least  sensitive  to  a  lack  of  vitamiue 
C.  Schaumann  (I.e.  2)  fed  dogs  on  denatured  meat  and  described 
a  condition  similar  to  beriberi,  with  paresis  and  changes  in  the 
pharynx  and  tongue ;  the  latter  appears  to  be  more  likely  of  a  scorbutic 
nature.  Despite  this,  Schaumann  (403)  held  that  all  these  symptoms 
could  be  removed  by  administering  yeast,  and  since  yeast  contains 
no  vitamine  C,  he  must  have  been  dealing  with  beriberi.  It  should 
be  pointed  out  here  that  the  findings  on  beriberi  in  dogs  are  not 
concordant.  Theiler,  Green  and  Viljoen  (I.e.  278)  stated  that  they 
could  not  produce  beriberi  or  scurvy  in  dogs  by  feeding  with  white 
rice;  the  animals  merely  suffered  from  malnutrition.  Voegtlin  and 
Lake  (I.e.  393),  on  the  contrary,  described  true  beriberi  in  dogs  fed 
on  meat,  extracted  with  soda. 

Karr  (404)  fed  dogs  on  lard,  cane  sugar,  salts  and  protein,  in  the 
form  of  casein  or  wheat  gluten.  After  3  to  9  weeks,  the  animals 
refused  the  food.  It  was  noted  that  grown  dogs  needed  very 
little  vitamines  A  and  C,  since  yeast  alone,  in  amounts  of  0.1 
gram  daily,  restored  the  appetite  to  normal,  while  an  addition  of  0.2 
gram  cured  beriberi  in  8  to  12  hours.  The  utilization  of  protein 
seemed  uninfluenced  by  the  presence  of  vitamine  B. 

Proceeding  to  the  rachitic-like  disease  in  dogs,  we  come  first  to 
the  work  of  Stilling  and  v.  Mering  (405)  in  1889,  who  fed  dogs  on 


138 


THE   VITAMINES 


extracted  meat  and  fat  in  order  to  study  osteomalacia  and  the  effect 
of  a  calcium-poor  diet.  The  experiment  lasted  126  days.  Hebrant 
•and  Antoine  (406)  described  a  typical  osteomalacia  in  dogs.  Guerin 
(407)  observed  rickets  in  young  dogs  fed  on  meat,  while  animals  from 
the  same  litter,  which  were  suckled,  remained  unaffected,  Some  of 
these  experiments  were  made  not  only  with  a  vitamine-poor  but  also 
with  a  calciurcnpoor  diet  and  the  pathological  condition  described 
was  nothing  but  osteoporosis.  To  this  category,  belongs  the  work 
of  Roloff  (407a)  in  1879,  and  also  the  communication  of  Reimers 
and  Boye  (407b).  Bull  (408)  described  a  severe  occurrence  of  rickets 
in  young  dogs  fed  on  cooked  meat  and  pollard,  in  Adelaide,  Australia. 
However,  the  systematic  investigations  of  Mellanby  (I.e.  95)  are 
of  chief  interest  in  this  chapter.  This  investigator  fed  young  dogs 
5  to  8  weeks  old  on  diets  of  various  composition,  poor  in  vitamine  A. 
These  diets  were  more  or  less  suitable  for  the  production  of  rickets 
and  in  the  end,  the  following  were  chosen  for  this  purpose. 


DIET  A 

DIET  B 

Skimmed  milk  .             

175  cc. 

250-350  cc. 

Wheat  bread  (white) 

ad  libitum 

ad  libitum 

Linseed  oil  

10  cc. 

10  cc. 

Yeast  

10  grams 

5-10  grams 

Orange  juice 

3  cc. 

NaCl  

1-2  grams 

1-2  grams 

Some  200  dogs  were  used  in  this  work;  at  first,  they  were  kept  on 
the  diet  for  6  months  but  later,  a  shorter  time,  for  it  was  demon- 
strated that  dogs  on  diet  B  developed  rachitic  symptoms  after  6 
weeks.  Mellanby  investigated  the  behavior  of  various  foodstuffs  of 
animal  and  vegetable  origin,  and  found  that  those  that  are  protective 
against  rickets  are  rich  in  vitamine  A.  Still,  meat  poor  in  fat  had 
some  effect  in  delaying  the  onset  of  rickets.  Quickly  growing  dogs 
develop  rickets  soonest,  which  apparently  shows  that  the  more 
active  the  growth  the  more  vitamine  A  is  necessary. 

Symptomatology.  The  ossification  of  the  bones  proceeds  in  an 
abnormal  manner;  hence  the  bones  bend  and  the  ligaments  become 
loose,  making  the  deformities  even  more  apparent.  Swelling  of  the 
epiphyses  of  the  bones  was  easily  noticeable  and  according  to  the 
severity  of  the  condition,  deformities  of  the  thorax  developed  with  a 


FIG.  33.  RICKETS  IN  A  DOG  (MELLANBY;  REP.  MED.  RES.  COM.) 


FIG.  34.  X-RAY  OF  BONE  OF  RACHITIC  PUP  (MELLANBY;  REP.  MED.  RES.  COM.) 

139 


140  THE   VITAMINES 

typical  rosary.  The  animals  were  very  little  resistant  to  infections, 
were  apathetic,  lethargic  and  extremely  unwilling  to  exert  them- 
selves, at  a  time  when  the  existing  bone  changes  could  not  yet  have 
been  responsible.  This  was  undoubtedly  due  to  the  impairment  of 
the  muscular  tonus. 

After  death,  the  bones  were  found  to  be  very  poor  in  calcium  and 
showed^  in  the  X-ray,  an  appearance  characteristic  of  rickets,  as 
may  be  seen  in  the  illustration.  Of  two  dogs  of  the  same  litter, 
one  animal  received  only  the  "rachitic"  diet,  the  other,  vitamine  A 
in  addition.  The  malformation  of  the  cartilage  epiphyses  and  the 
more  frequently  occurring  osteoid  tissue  are  the  two  chief  symptoms. 
According  to  Henderson  (408c)  the  muscle  of  the  rachitic  dog  con- 
tains less  creatine  than  the  normal.  Findlay,  Paton  and  Sharpe 
(408b)  conducted  metabolism  experiments  on  rachitic  dogs.  A  cal- 
cium-poor diet  caused  osteoporosis,  but  not  rickets,  while  in  rachitic 
dogs  the  blood  and  the  tissues  showed  the  normal  calcium  content. 

The  significance  of  Mellanby's  experiments  for  the  study  of  rickets 
will  be  discussed  further  in  the  description  of  this  disease. 

In  conclusion,  we  shall  mention  an  investigation  of  Chittenden  and 
Underhill  (I.e.  28).  They  fed  dogs  on  cooked  peas,  zwieback  and 
cottonseed  oil,  and  noted  a  disease  which  they  believed  to  be  anal- 
ogous to  pellagra  in  man.  The  disease  developed  after  2  to  8  months, 
showing  that  a  larger  amount  of  peas  was  less  harmful.  The  dis- 
ease, which  could  be  cured  by  adding  meat  to  the  diet,  produced 
ulcerations  of  the  mouth;  in  particular,  the  entire  enteron  was 
hemorrhagic.  Besides,  there  was  observed  a  great  loss  of  weight, 
peculiar  gait  and  diarrhoea.  If  a  part  of  the  peas  was  substituted 
by  casein,  the  symptoms  appeared  sooner.  This  shows,  perhaps, 
that  the  condition  is  associated  with  a  lack  of  vitamines,  since  peas 
contain  some  vitamine  but  not  enough.  To  our  knowledge,  this  work 
has  not  yet  been  repeated  by  any  other  investigator. 

Domestic  animals 

We  come  now  to  the  consideration  of  domestic  animals,  among 
which  we  shall  discuss  goats,  sheep,  horses  and  pigs.  In  this  field, 
there  are  questions  which  are  not  yet  quite  clear,  but  which  possess  a 
definite  significance  in  their  social  and  economic  aspect.  We  have 
very  little  accurate  knowledge  of  the  vitamine  requirements  of  these 
animals.  In  various  parts  of  the  world,  we  encounter  diseases, 


THE   VITAMINE    IN   THE    ANIMAL   KINGDOMS  141 

which,  though  not  yet  explained  etiologically,  nevertheless  always 
give  some  indications  showing  that  they  may  be  avitaminoses.  In 
most  cases,  they  occur  on  a  large  scale  in  Nature,  and  consequently 
are  specially  deserving  of  our  interest. 

Sheep  and  goats.  Experimental  investigations  with  these  animals 
were  first  made  by  Schaumann  (I.e.  2).  A  goat,  fed  on  corn  and 
white  rice,  lost  weight  rapidly  and  showed,  according  to  Schaumann, 
paresis,  although  corn  certainly  contains  sufficient  vitamine  B.  The 
animal  was  then  given  yeast,  Katjang-idjoe  beans,  and  green  fodder 
(the  latter  for  one  month),  with  a  resultant  gain  in  weight.  This 
effect  was  attributed  to  the  yeast  and  the  beans,  although  the  green 
fodder  very  likely  played  a  greater  role.  This  experiment  was  not 
clear  and  consequently  is  of  little  significance.  Henriques  and 
Andersen  (409)  administered  parenterally  hydrolyzed  meat,  glucose, 
sodium  acetate  and  salts  to  goats.  For  20  days,  a  marked  nitrogen 
retention  was  noted.  They  then  heated  the  food  mixture  for  20 
minutes  at  110°C.,  and  were  able  to  show  that  it  was  no  longer 
possible  to  maintain  the  nitrogen  balance,  except  when  the  heating 
was  not  carried  beyond  100°C.  These  results  were  attributed  to  the 
susceptibility  of  tryptophane  to  heat.  Fingerling  (410)  fed  lambs  on 
inorganic  and  organic  phosphorus  combinations,  without  observing 
any  particular  differences.  He  made  the  correct  observation  that 
the  frequently  noted  improper  nutrition  with  inorganic  phosphorus 
compounds  was  not  due  to  the  character  of  the  phosphorus,  but  to 
the  composition  of  the  food  itself. 

Theiler,  Green  and  Viljoen  (I.e.  278)  were  unable  to  demonstrate 
an  avitaminosis  in  sheep  and  goats.  These  animals  tolerated  the 
rice  diet  better  when  an  addition  of  autoclaved  hay  or  straw  was 
made.  Very  soon,  enteritis  made  its  appearance,  though  in  the 
opinion  of  these  investigators,  this  had  nothing  to  do  with  the  diet. 
At  all  events,  the  requirements  of  vitamine  B  appear  to  be  very 
modest. 

The  naturally  occurring  nutritional  disturbances  were  described  by 
Hoare  (411)  as  a  form  of  edema,  which  develops  because  of  an  insuffi- 
cient fodder  and  poor  climatic  conditions.  A  disease  of  sheep,  known 
as  "staggers,"  was  described  by  Jones  and  Arnold  (412)  in  South 
America.  It  is  a  nervous  disturbance  of  sheep,  as  well  as  horses  and 
cattle,  caused  by  the  consumption  of  coarse  grass  (Poa  Argentina). 
They  were  not  clear  as  to  the  etiology,  and  were  more  inclined 


142  THE   VITAMINES 

• 

towards  the  conception  of  a  toxic  phenomenon.  The  observed 
symptoms  were  general  weakness,  muscle  tremors,  peculiar  movement 
of  the  head,  stiffness  of  the  limbs,  intermittent  paralysis  with  spastic 
appearances,  particularly  after  irritation,  impairment  of  the  visual 
capacity  and  conjunctivitis.  The  disease  could  be  produced  experi- 
mentally by  feeding  pampas  grass.  The  time  necessary  for  this 
varied  between  2  and  21  days,  averaging  10  days,  during  which 
spontaneous  cures  often  occurred.  By  changing  the  diet,  it  was 
possible  to  effect  a  cure.  Young  animals  were  far  more  susceptible 
than  grown  ones,  and  the  disease  was  attributed  to  a  poison  in  the 
grass,  present  at  all  seasons  and  in  all  parts  of  plants.  The  post- 
mortem revealed  nothing  positive  and  afforded  no  definite  indications 
of  the  nature  of  the  toxin.  Reid  and  Aston  (413)  observed  a  bone 
cachexia  in  sheep,  which  they  referred  back  to  a  lack  of  inorganic 
factors  of  the  diet. 

Reindeer.  Ibele  (414)  noted  a  disease  called  "Lecksucht"  supposed 
to  be  due  to  the  lack  of  alkaline  earths. 

It  is  difficult,  without  having  personally  seen  these  animals,  to 
draw  a  picture  of  the  nature  of  the  disease.  The  observation  that 
in  one  case  the  disease  could  be  produced  after  two  days  feeding, 
would  speak  against  its  being  an  avitaminosis,  if  the  investigators 
had  not  explicitly  emphasized  that  this  is  possible  only  in  certain 
susceptible  animals.  This  lack  of  resistance  can  be  explained,  at 
least  theoretically,  by  a  previously  occurring  deficient  dietary. 

Pigs.  The  nutrition  of  these  animals  was  studied  from  two  view- 
points— first,  from  that  of  the  animal  breeders  in  connection  with  the 
food  requirements,  and  secondly,  from  that  of  laboratory  men  in 
relation  to  the  vitamines.  It  is  true,  as  Evvard  (415)  pointed  out, 
that  if  these  animals  are  permitted  to  select  their  own  food,  their 
natural  instinct  will  lead  them  to  choose  a  correct  diet.  Conditions 
are  different  in  winter,  however,  when  a  prepared  diet  must  be 
provided.  Hart  and  McCollum,  as  well  as  their  co-workers,  studied 
the  exact  food  requirements  of  pigs  in  relation  to  growth,  maintenance 
and  reproduction.  Such  studies  were  published  by  Hart,  McCollum 
and  Fuller  (416)  in  1908.  They  showed  that  pigs  fed  on  rice  and 
washed  bran  suffered  from  osteoporosis,  while  those  fed  on  unwashed 
bran  did  not  manifest  this  disease.  At  that  time,  the  conclusion 
was  made  that  the  phosphorus  compounds  were  washed  out  with 
water — which  of  course  does  not  hold  today.  Hart  and  McCollum 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  143 

(417)  observed  that  between  wheat  and  corn,  there  are  great  differ- 
ences as  regards  the  growth-promoting  properties.  While  the 
animals  attained  a  weight  of  100  pounds  on  wheat,  on  corn  they 
weighed  from  200  to  300  pounds.  McCollum  (418)  fed  young  pigs 
on  wheat,  oats  and  corn  and  found  that  casein,  as  well  as  skimmed 
milk,  was  a  better  source  of  nitrogen.  After  three  months,  the 
animals  declined  and  McCollum  left  the  question  open  as  to  whether 
this  condition  was  due  to  lack  of  salts  or  vitamine,  or  to  the  presence 
of  a  .toxic  substance.  Hart  and  McCollum  (419)  could  not  keep 
young  pigs  alive  for  a  long  time  on  corn  and  gluten,  while  the  addition 
of  casein  or  milk  appreciably  improved  growth.  Hart  and  Steenbock 
(420)  investigated  a  series  of  plant  products,  with  the  addition  of 
the  same  animal  foods  as  above,  and  came  to  the  conclusion  that 
rice,  wheat,  corn,  potatoes,  and  cabbage  possessed  little  nutritive 
value,  but  it  was  not  further  investigated  whether  the  vitamines 
played  a  role.  Later,  Hart  and  Steenbock  (421)  sought  to  settle 
the  question  as  to  whether  the  presence  or  absence  of  vitamines  has 
any  effect.  For  this  purpose,  grown  pigs  were  kept  for  a  long  time 
on  corn  and  oats.  Eventually,  symptoms  of  the  disease  appeared, 
such  as  stiffness  of  the  legs  and  difficulty  of  getting  about,  these 
conditions  being  specially  manifested  in  pregnancy  and  lactation. 
A  great  number  of  young  were  born  dead.  If  these  symptoms  were 
not  noticed  in  the  first  litter,  they  were  present  in  the  second.  In 
most  cases,  an  improvement  was  seen  when  the  cereal  diet  was  sub- 
stituted by  a  corn-alfalfa  mixture,  which  is  much  richer  in  vitamines. 
The  dietary  deficiency  was  however  attributed  largely  to  the  unfavor- 
able composition  of  the  inorganic  constituents.  These  exact  experi- 
ments are  particularly  important  for  the  recognition  of  the  etiology 
of  some  very  frequently  occurring  diseases  of  cattle  which  are  of 
practical  importance,  and  which  have  lately  been  quite  often  attrib- 
uted to  the  effect  of  a  poison.  We  see,  however,  that  feeding  in 
the  stable  may  lead  to  dietary  deficiencies,  without  it  being  necessary 
to  assume  a  toxic  cause. 

The  superiority  of  skimmed  milk  over  all  other  food  mixtures  was 
also  set  forth  in  a  number  of  publications  of  Klein  (422).  This 
favorable  influence  may  be  explained  either  by  a  better  utilization  of 
the  protein,  by  the  vitamine  content,  or  by  both  factors. 

Despite  all  these  investigations,  and  still  others  to  be  mentioned, 
we  are  not  yet  accurately  informed  as  to  the  vitamine  requirements  of 


144  THE   VITAMINES 

pigs.  This  is  so  because  few  investigations  in  which  only  the  vita- 
mine  factor  is  lacking,  while  all  other  factors  are  controlled,  have  been 
carried  out.  All  of  the  above-mentioned  investigations  are  com- 
plicated by  mixed  symptoms,  and  it  is  therefore  impossible  to  form 
an  unbiased  opinion  as  to  the  nutritive  requirements  of  these  animals. 
This  is  all  the  more  regrettable,  since  the  pig,  which  eats  anything, 
is  peculiarly  adapted  for  the  study  of  human  nutrition. 

In  1907,  Hoist  and  Frolich  (I.e.  36)  described,  in  rice-fed  pigs, 
symptoms  which  were  thought  to  be  mixed  forms  of  beriberi  and 
scurvy.  Since  white  rice  is  lacking  in  all  three  vitamines,  it  is  impos- 
sible to  say  which  one  is  most  important  for  these  animals.  Theiler, 
Green  and  Viljoen  (I.e.  278)  on  the  contrary,  could  observe  no 
specific  symptoms  on  this  food.  In  one  case,  gall  stones  were  found; 
in  others,  enteritis  was  noted,  which  may  perhaps  be  regarded  as  a 
sign  of  an  avitaminosis.  Plimmer  (422a)  observed  scurvy  in  pigs 
fed  on  cooked  food,  the  symptoms  of  which  were  relieved  by  the 
administration  of  uncooked  food.  Green  (423)  carried  out  investiga- 
tions with  a  diet  poor  in  calcium,  and  in  this  way  found  two  kinds 
of  undernourishment;  one  kind  depends  upon  protein  which  was 
qualitatively  shown  to  be  inadequate,  while  the  second  could  be 
attributed  to  insufficient  protein  and  salts  addition.  The  animals 
were  in  poor  nutritive  condition  and  showed  defective  bone  growth 
with  insufficient  calcification.  Substitution  of  lard  by  butter  exerted 
no  favorable  influence.  It  is  one  of  the  few  investigations  on  the 
nutrition  of  domestic  animals,  in  which  vitamine  A  was  given  special 
attention.  Another  investigation,  in  which  the  presence  of  vitamine 
A  in  the  diet  was  provided  for,  was  that  of  Hart,  Miller  and  McCollum 
(424)  A  typical  composition  of  the  diet  used  by  them  was  95.5 
parts  wheat  meal,  2.5  parts  wheat  gluten,  2  parts  butter,  di-potassium 
phosphate  and  calcium  lactate.  On  this  diet,  the  animals  developed 
symptoms  quite  similar  to  those  of  beriberi.  Among  other  symp- 
toms, pathological  changes  were  noted  in  the  spinal  cord  which 
showed  fatty  degeneration.  The  symptoms  made  their  appearance 
afte,r  about  9  months,  when  a  decrease  in  weight,  difficulty  of  forward 
motion,  rough  coat,  labored  breathing  and  muscle  tremors  were 
observed.  There  was  also  stiffness  of  the  extremities  and  dragging 
of  the  legs;  if  the  animals  were  helped  to  their  legs  they  again  sank 
at  the  knees.  Since  McCollum,  at  this  time,  denied  the  existence 
of  vitamine  C,  the  diet  used  was  considered  as  complete,  and  the 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  145 

disease  was  attributed  to  toxic  action  of  wheat.  Addition  of  whole 
milk  had  only  a  slight  effect  upon  the  general  well-being  of  the 
animals,  but  the  addition  of  alfalfa  grass  soon  permitted  of  the 
development  of  a  normal  condition.  In  all  these  investigations, 
growth  was  interrupted  when  the  general  condition  of  the  animal 
became  worse.  It  seems  to  us  that  without  further  indisputable 
evidence,  it  is  difficult  to  accept  the  view  as  to  the  toxicity  of  wheat 
kernel.  It  may  be  possible  that  food  mixtures  were  used,  the  vita- 


FIG.  35.  BERIBERI-LIKE  DISEASE  IN  A  PIG  ON  A  DIET  CONSISTING  OF  45  PARTS 

CORN,  45  PARTS  OATS,  10  PARTS  OIL  SEED  PRESS-CAKE  AND  5  PARTS 

BUTTER  (HART,  MILLER  AND  MCCOLLUM) 

mine  A  content  of  which  was  barely  sufficient,  while  vitamine  C  was 
almost  completely  lacking.  Experiments  with  the  addition  of  these 
vitamines  were  not  made. 

With  products  derived  from  cotton  seed,  the  behavior  was  naturally 
different.  These  contain  a  toxic  substance  called  "gossypol."  Such 
products,  fed  to  pigs,  cause  death  in  50  to  80  days,  according  to 
Roberts  (425) .  But  even  in  these  cases,  there  seems  to  be  a  vitamine 
deficiency,  according  to  Rommel  and  Vedder  (426).  In  8  to  15  days, 
this  food  produces  in  these  animals  a  picture  of  disease  resembling 


146  THE   VITAMINES 

perhaps  the  wet  form  of  beriberi  in  man,  and  quite  analogous  to  the 
symptoms  which  pigs  develop  when  fed  white  rice.  Zilva  and  col- 
laborators (426a)  found  that  the  pig  was  susceptible  to  the  develop- 
ment of  experimental  rickets,  but  in  this  animal,  the  lack  of  vitamine 
A  (of  milk)  was  not  the  sole  causative  factor. 

In  spite  of  this  series  of  results  we  are  not  yet  clear  as  to  the  vita- 
mine  requirements  of  pigs,  so  that  further  work  is  necessary. 

Horses.  In  1908,  Friedberger  and  Froehner  (427)  described  edema 
in  horses  and  oxen  which  performed  heavy  work  in  the  sugar  refiner- 
ies, and  were  fed  on  sugar  beet  residues.  These  residues  contained 
only  a  slight  amount  of  protein,  about  0.5  per  cent,  very  little  dry 
substance  (5  per  cent)  and  high  water  content  (95  per  cent).  This 
diet  which,  aside  from  the  protein,  appeared  to  be  lacking  in  other 
constituents,  together  with  the  hard  work  and  high  water  content  of 
the  food,  was  supposed  to  be  responsible  for  the  development  of  this 
disease.  Similar  pathological  conditions  were  likewise  noted  in 
horses  by  Hutyra  and  Marek  (428). 

Scheunert,  Schattke  and  Lotsch  (429)  observed  a  disease  in  horses, 
resembling  osteomalacia,  in  the  poor  mountainous  districts  in  Saxony. 
Kawakami  (430)  observed  in  horses,  a  disease  called  "sukumi"  or 
"gokusukumi,"  which  developed  when  rice,  barley  and  oats  were  fed. 
The  disease  exhibits  a  certain  similarity  to  experimental  beriberi  and 
starts  with  gastro-intestinal  disturbances.  Theiler,  Green  and 
Viljoen  (I.e.  278)  fed  horses  on  white  rice  for  six  months  without 
noticing  any  signs  of  an  avitaminosis.  This  is  the  more  remarkable 
since  white  rice  is  supposed  to  be  lacking  in  protein,  salts  and  vitamine 
A,  besides  vitamine  B. 

Cows  and  oxen.  In  1913,  the  late  Dr.  Donald  Macauley  called  our 
attention  to  a  South  African  cattle  disease,  known  there  as  "Stijf- 
ziekte"  and  "Lamziekte."  On  perusing  the  available  literature,  it  is 
apparent  that  similar  conditions  are  known  in  all  parts  of  'the  world. 
Stewart  (431),  for  instance,  reported  a  disease  known  in  Australia 
mostly  as  "rickets."16  In  the  poor  mountainous  districts  of  Sax- 
ony, according  to  the  report  of  Lotsch  (432),  there  is  not  so  infre- 
quently a  pathological  condition  called  "Stallm angel,"  supposed  to 
be  caused  by  a  diet  poor  in  salts.  This  severe  disturbance  of  metabo- 
lism reminds  one  strongly  of  human  rickets  and  osteomalacia. 

16  Stewart  was  disposed  to  consider  the  disease  as  an  avitaminosis. 


THE   VITAMINES   IN   THE    ANIMAL   KINGDOM  147 

From  New  Zealand,  there  was  a  report  by  Aston  and  Reakes  (433) 
on  progressive  anemia,  known  locally  as  "bush-sickness."  It  was 
supposed  to  be  caused  by  lack  of  salts,  and  differed  from  lamziekte. 
In  addition  to  these  naturally  occurring  pathological  conditions,  we 
have  the  experience  gained  by  the  experimental  feeding  of  cattle. 
Russel  and  Morrison  (434)  found  that  cattle  fed  on  oats,  straw  and 
butter,  or  casein,  butter  and  oat-straw,  gave  birth  to  very  miserable 
looking  offspring.  This  condition  was  not  regarded  as  an  avitam- 
inosis  but  as  a  lack  of  calcium,  since  an  addition  of  these  salts  seemed 
to  improve  the  condition  of  the  animals.  Fleischmann  (435)  noted 
that  calves  fed  on  hay  often  became  sick,  and  he  proceeded  to  study 
the  chemical  changes  attending  the  drying  of  the  grass.  He  showed 
that  lecithin  and  phosphoproteid,  as  well  as  protein,  underwent 
some  decomposition  during  the  process  of  drying.  However,  since 
the  total  nitrogen  showed  no  change,  we  may  assume  that  there  was 
no  loss  of  inorganic  constituents.  The  real  difference  observed  must 
be  looked  for  in  the  vitamine  content. 

We  find  a  somewhat  different  conception  in  the  work  of  Henry 
(436).  In  the  description  of  an  Australian  cattle  disease  known  as- 
"impaction  paralysis,"  resembling  lamziekte,  he  attributes  the  cause 
of  the  disease  to  the  poverty  of  the  soil  in  calcium  and  phosphorus. 
The  disease  appears  at  the  end  of  the  dry  season,  in  places  where  the 
vegetation  is  injured  by  the  Australian  rabbit-plague.  The  disease 
was  found  where  there  was  an  abnormal  desire  to  eat  bones  and 
where  osteomalacia  had  also  prevailed  for  some  time.  A  favorable 
effect  was  noted  on  adding  a  nourishing  diet,  such  as  skimmed  milk. 
We  have  already  mentioned  the  interesting  assumption  of  Davis 
(I.e.  195),  who  held  that  in  the  last  analysis  the  poverty  of  the  soil 
was  the  cause  of  diseases  of  domestic  animals.  When  the  vegetation 
is  so  impoverished  through  extraneous  conditions  that  it  no  longer 
provides  sufficient  calcium  and  phosphorus  for  the  animals,  it  is- 
conceivable  that  other  important  substances,  for  example,  vitamines, 
might  also  be  lacking.  It  must  likewise  not  be  forgotten  that  in  the 
avitaminoses  there  is  frequently  a  loss  of  inorganic  constituents, 
and  this  condition  might  manifest  itself  in  an  abnormal  desire  for 
bones  or  other  materials  rich  in  calcium  and  phosphorus.  For 
example,  Forbes  (437)  stated  that  domestic  animals  often  display  an 
abnormal  craving  for  mineral  substances.  Pigs  and  cattle  sometimes 
eat  large  quantities  of  bone  meal,  with  resultant  improvement  in 


148 


THE   VITAMINES 


r 


health.  Cattle  growers  know  that  the  condition  of  the  legs  in  horses 
depends  upon  the  fodder  and  largely  on  the  nature  of  the  soil.  Place 
(438)  believed  he  saw  among  cattle  not  only  diseases  similar  to  beri- 
beri but  also  scurvy-like  conditions  in  cattle,  horses  and  sheep, 
infantile  scurvy  in  calves  and  sheep,  and  diseases  of  the  type  of 
pellagra,  with  skin  and  gastro-intestinal  symptoms. 

Stijfziekte  and  lamziekte.  These  diseases  of  cattle  and  goats  occur 
widely  in  South  Africa,  and  are  described  by  Theiler  (439).  These 
conditions,  particularly  lamziekte,  have  become  of  such  economic 
importance  that  a  great  number  of  well-known  investigators  have 

occupied  themselves  with  this  problem. 
In  some  severely  affected  districts,  the 
lamziekte  problem  has  become  so  acute 
that  it  no  longer  pays  to  breed  cattle 
because  of  the  great  mortality.  Of 
the  two  diseases,  stijfziekte  has  been 
least  investigated,  apparently  because 
of  its  lesser  practical  importance.  It 
is  a  bone  disease  of  young  animals, 
accompanied  primarily  by  swelling  of 
the  joints,  the  metatarsals  and  meta- 
carpals,  and  secondarily,  the  epiphyses 
of  the  long  bones.  These  swellings  are 
painful  and  since  the  fore-legs  are  most 
affected,  the  animal  tries,  as  far  as 
possible,  not  to  use  them,  and  runs 
around  with  forward-bent  hind  legs 
and  hunched  back.  Muscular  weak- 
ness and  abnormal  appetite  (pica)  are 
among  some  of  the  symptoms  of  the  disease.  The  temperature 
remains  normal.  The  diaphyses  of  the  diseased  bones,  on  sectioning, 
appear  deep  red  and  blotched  with  bloody-serous  liquid. 

Some  forms  of  stijfziekte  resemble  lamziekte  very  much,  without 
necessarily  indicating  a  relationship  between  the  two,  in  the  opinion 
of  Theiler,  Green  and  Viljoen  (I.e.  278).  Theiler  admitted  that 
stijfziekte  might  be  an  avitaminosis.  If  this  were  so,  it  would  help 
clear  up  the  problem  of  lamziekte,  since  it  could  serve  as  a  proof  that 
the  nourishment  of  cattle  in  some  sections  of  South  Africa  could  be 
improved.  As  it  happens,  stijfziekte  may  be  cured  by  a  change  of 


FIG.  36.  STIJFZIEKTE  (THEILER) 


THE   VITAMINES   IN   THE   ANIMAL   KINGDOM  149 

diet,  according  to  available  data.  Feeding  of  good  grass,  green 
barley  and  green  millet  is  supposed  to  be  accompanied  by  beneficial 
results. 

Lamziekte  or  gal-lamziekte.  The  history  of  lamziekte  research  is 
particularly  instructive  since  it  shows  us  how  easy  it  is  to  go  from  one 
extreme  to  another.  In  times  of  great  popularity  of  the  vitamine 
theory,  one  is  tempted  to  classify  every  condition  that  bears  some 
analogy  to  the  already  known  avitaminoses  in  the  same  group. 
Lamziekte,  according  to  the  present  stage  of  investigation  at  least, 
is  not  an  avitaminosis  and  would  not  have  found  a  place  in  our 
treatise  if  it  were  not  for  the  possibility  of  explaining  one  of  the 
etiological  factors  through  a  lack  of  vitamine. 

This  disease  is  prevalent  in  all  breeds  of  cattle  without  regard  to 
species  or  sex.  Young,  pregnant  or  milking  animals  appear  to  be 
specially  susceptible.  Lamziekte  occurs  particularly  after  long 
periods  of  draught,  and  is  manifested  by  paralysis  and  contractions 
associated  with  the  degeneration  of  the  peripheral  and  central 
nervous  system,  roughly  resembling  avian  beriberi.  We  note  here 
paralysis  of  the  legs,  opisthotonos,  dysphagia  and  paresis  of  the 
tongue.  Lack  of  appetite  and  pica  (abnormal  craving),  excessive 
salivation,  loss  of  milk,  constipation  and  also  diarrhoea — all  these 
constitute  the  earlier  symptoms.  The  temperature  appears  to  be 
subnormal.  The  usual  progress  is  sub-acute  and  lasts  some  weeks; 
the  animals  sometimes  show  an  apparent  improvement,  but  fatal 
results  ensue  when  a  second  attack  occurs.  In  spite  of  a  cure,  the 
contractions  may  still  persist  for  a  long  time.  Acute  forms  last 
from  two  to  seven  days,  and  yet  sudden  developments  are  noted,  in 
which  apparently  healthy  animals  fall  into  a  comatose  condition  in 
10  to  20  hours,  which,  accompanied  by  sub-normal  temperature, 
results  in  death.  This  condition  has  been  frequently  confused  by 
South  African  farmers  with  another  called  "poverty,"  and  resembling 
malnutrition.  This  condition,  "poverty,"  may  eventually  prove  to 
be  a  concealed  avitaminosis. 

Pathological  anatomy  of  lamziekte.  The  findings  have  been  spe- 
cially described  by  Hedinger  (440).  They  consist  of  hydrothorax, 
and  ecchymoses  of  the  pleura,  epicardium,  endocardium  and  thymus; 
hyperemia  was  observed  in  all  the  abdominal  organs,  in  the  mucosa 
of  the  fourth  stomach,  intestines  and  lungs;  a  moderate  hydroperi- 
cardium  was  frequently  noted;  enteritis  of  the  small  intestine, 
coupled  with  hemorrhages  and  ascites,  with  a  dilatation  of  the  heart, 


150  THE   VITAMINES 

occurred  not  infrequently.     In  the  bones,  on  the  contrary,  at  least 
microscopically,  no  changes  were  found. 

Etiology.  We  may  well  imagine  that  because  of  the  great  practical 
significance  of  this  disease  no  effort  has  been  spared  to  discover  its 
cause.  One  of  the  older  theories  of  Theiler  (I.e.  439)  assumed  the 
presence  of  toxins  in  the  grass.  This  theory  was  experimentally 
tested  by  Viljoen  (441)  in  that  he  fed  a  series  of  animals  on 
various  kinds  of  grass,  found  in  regions  affected  by  lamziekte,  with 
negative  results.  The  theory  of  infection  was  tested  by  Spruell 
(442);  cows  and  goats  received  intravenous  injections  of  the  blood, 
lymph  glands  and  ascites  fluid  of  sick  animals;  here  too,  the  experi- 
ments proved  to  be  negative.  Mitchell  (443)  and  Walker  (444) 
sought  to  bring  about  the  disease  in  healthy  animals  by  using  carcass 
material  of  lamziekte  animals,  but  in  vain.  We  formerly  held  that 
lamziekte  was  similar  to  beriberi,  but  this  view  was  modified  by  us 
in  1915  (445),  in  that  we  considered  it  to  be  similar  to  rickets. 
This  view  is  also  held  by  Hart,  Steenbock  and  Hoppert  (445a),  who 
arrived  at  this  conclusion  from  experiments  with  goats.  Here  cod 
liver  oil  had  a  distinctly  favorable  action  on  the  calcium  metabolism. 
In  spite  of  this,  our  first  opinion  was  carefully  tested,  and  Stead  (446) 
attempted  to  show  that  lamziekte  was  analogous  to  beriberi.  He  fed 
pigs  on  meat  obtained  from  animals  which  had  died  of  lamziekte,  in 
the  belief  that  it  was  poorer  in  vitamine  B  than  normal  meat. 
Cattle  were  also  fed  on  white  rice,  samp  (a  product  made  from 
decorticated  corn),  white  flour  and  oats;  they  showed  pica,  stiffness 
of  the  legs  and  pains  on  walking.  Theiler,  Green  and  Viljoen  (I.e. 
278)  have  investigated  a  series  of  animal  species,  already  mentioned 
previously,  as  to  their  vitamine  B  requirements;  horses,  dogs,  goats, 
sheep  and  pigs  were  used.  They  were  kept  on  a  diet  consisting  chiefly 
of  white  rice,  and  in  no  case  was  there  any  symptom  characteristic 
of  beriberi.  Cattle  were  kept  on  white  rice  for  a  year  without  the 
appearance  of  any  beriberi  symptoms.  In  the  worst  case,  symptoms 
were  noted,  in  accord  with  the  findings  of  Stead,  which  were  thought 
of  as  laminitis  and  which  could  not  be  attributed  to  a  lack  of  vitamine 
B.  Latterly,  Stead  seems  to  have  abandoned  his  view  in  favor  of 
Theiler's.17  One  cow  fed  on  white  rice  and  autoclaved  straw  or 

17  Theiler  said  that  no  grass  could  be  so  poor  in  vitamine  as  was  the  diet 
which  he  used  and  he  believed,  therefore,  that  it  had  nothing  to  do  with  an 
avitaminosis.  The  survival  of  the  animal,  he  attributed  to  a  bacterial 
symbiosis  (p.  75). 


THE   VITAMINES   IN   THE    ANIMAL   KINGDOM  151 

hay,  gave  birth  to  a  calf  that  was  blind.  The  validity  of  the  vitamine 
hypothesis  was  tested  in  two  ways.  First,  through  a  repetition  of 
Stead's  experiments,  by  feeding  to  healthy  animals  the  meat  of 
animals  which  had  died  of  lamziekte,  definite  indication  as  regards 
impoverishment  of  vitamine  B  could  not  be  found.  Secondly,  the 
influence  of  substances  rich  in  vitamine  B  was  studied.  An  addition 
of  orypan  (extract  of  rice  polishings)  and  yeast  had  no  effect,  and 
we  may  therefore  assume,  with  certainty,  that  lamziekte  cannot  be 
caused  by  a  lack  of  vitamine  B.  Experiments  with  other  vitamines 
were  not  tried. 

In  the  meantime,  Theiler  and  his  co-workers  (447)  have  apparently 
completely  cleared  up  the  etiology  of  lamziekte.  They  maintain  that 
in  the  etiology  of  this  disease,  six  phases  prevail,  which  are  responsible 
for  its  occurrence,  and  are  related  to  each  other,  like  the  links  of  a 
chain. 

1.  A  toxin  or  poison  which  causes  the  disease. 

2.  Toxin-forming  saprophytes. 

3.  The  availability  of  carcasses  from  which  the  toxins  are  formed. 

4.  Pica,  or  abnormal  craving,  which  leads  animals  to  eat  carcasses, 
not  present  under  normal  conditions. 

5.  The  character  of  the  vegetation  and  the  soil  (and  climate)  upon 
which  pica  depends. 

6.  Susceptibility  of  the  animals  to  the  toxin. 

The  toxin  was  systematically  investigated  and  seemed  to  be 
similar  to  the  botulinus  toxin.  It  is  very  active,  inasmuch  as  0.00001 
cc.  per  kilo  body  weight  is  a  fatal  dose.  Lamziekte  may  also  be 
produced  experimentally  by  administration  of  this  toxin.  The  pro- 
phylaxis and  therapy  consist  of  the  elimination  of  one  of  the  six 
above  mentioned  links  of  the  chain,  at  least  theoretically.  Practi- 
cally, however,  it  seems  that  the  best  starting  point  is  the  pica  and 
since  this  condition  is  of  greatest  interest  to  us,  we  shall  speak  of 
it  in  greater  detail. 

Pica.  It  has  been  demonstrated  in  South  Africa,  that  not  only 
cattle  may  exhibit  this  condition,  but  also  ostriches,  goats  and 
poultry.  Pica  is  supposed  to  be  a  nervous  ailment,  which  arises 
only  where  the  condition  of  the  soil  is  such  that  the  water  quickly 
passes  through  the  upper  layers.  Because  of  this,  the  soil  becomes 
poor  in  phosphorus  and  hence  produces  a  vegetation  poor  in  this 
element.  Pica  itself  is  not  dangerous,  but  becomes  so  when  the 


152  THE   VITAMINES 

animals  find  infected  carcasses  to  eat.  This  condition  is  tested  by 
giving  the  animals  rotten  bones;  if  they  chew  these  bones,  then  the 
diagnosis  of  pica  is  verified. 

Cure  of  pica.  To  overcome  pica,  a  mixed  diet  may  be  used  which 
will  cure  the  disease  in  one  month.  Two  pounds  of  wheat  bran  cures 
it  in  three  weeks,  and  112  grams  of  bone  meal  is  supposed  to  have 
the  same  effect.  Besides  this,  the  addition  of  calcium  phosphate 
and  sodium  phosphate,  and  even  phosphoric  acid  in  drinking  water, 
may  be  used.18 

Etiology  of  pica.  As  we  have  seen  above,  Theiler  thought  that 
the  chief  cause  of  pica  was  the  poverty  of  the  diet  in  phosphorus, 
accompanied  perhaps  by  too  much  calcium,  although  it  is  admitted 
that  other  dietetic  factors  also  may  play  a  role.  Theiler  also  stated 
that  on  analysis  of  the  diet,  the  phosphorus  was  found  within  the 
limits  of  the  accepted  standards  for  such  animals.  He  concluded 
therefore,  that  the  accepted  figures  had  to  be  revised.  We  know 
from  the  study  of  deficiency  diseases,  that  under  such  conditions 
there  is  often  a  negative  calcium  and  phosphorus  balance,  and  it  has 
been  stated  from  time  to  time,  that  the  addition  of  phosphorus  (we 
refer  only  to  phospho-cod  liver  oil,  hypophosphites,  etc.)  may  result 
in  a  definite  improvement.  This  may  be  the  case  with  pica.  The 
animal  perhaps  has  sufficient  phosphorus  in  the  food  but  is  unable  to 
retain  it  because,  we  may  say,  the  diet  lacks  vitamine  A.  An  addition 
of  phosphorus,  in  whatever  form,  may  perhaps  lead  to  a  temporary 
improvement  and,  in  the  light  of  the  modern  conceptions  of  lamziekte, 
may  also  protect  against  this  disease,  since  the  animals  would  not 
eat  the  carcasses.  But  the  cause  of  pica  and  the  metabolic  disease 
may  endure  till  the  arrival  of  a  better  season  and  a  recovery  of  the 
vegetation.  For  it  can  be  shown  definitely  that  pica  prevails  only 
at  certain  times  of  the  year,  when  the  vegetation  thrives  but  poorly. 
If  this  is  so,  then  vitamines,  as  well  as  salts,  may  be  lacking;  a  normal 
vegetation  should  provide  everything  for  the  animal  that  is  necessary 
to  life.  The  simultaneous  development  of  undernutrition,  in  the 
form  of  "poverty,"  and  perhaps  also  stijfziekte,  in  lamziekte  regions, 
is  illustrative  of  the  profound  dietetic  deficiency  of  the  local  vegeta- 
tion. As  to  the  etiology  of  pica,  this  can  be  cleared  up  only  by  exact 
investigation,  especially  by  metabolism  experiments.  Such  accurate 

18  In  this  connection  it  is  interesting  to  compare  the  influence  of  phos- 
phorus on  pica  with  that  on  experimental  rickets  in  rats  (p.  327). 


THE   VITAMINES   IN  THE   ANIMAL   KINGDOM  153 

investigations  appear  still  to  be  lacking.  Above  all,  it  must  be 
determined,  in  the  event  that  the  disturbance  of  metabolism  may  be 
cured  by  the  addition  of  phosphorus,  how  vitamine  A  and  C,  which 
may  well  be  present  in  poor  vegetation  in  sub-optimal  amounts, 
would  act.  The  influence  of  these  vitamines  on  pica  has  been  very 
little  investigated. 

Monkeys 

Only  the  lower  types  of  monkey  have  been  used  of  late  for  vitamine 
studies;  the  first  work  was  that  of  Schaumann  (I.e.  2)  who  fed 
monkeys  on  washed  white  rice,  with  the  result  that  the  animals  died 
in  74  days.  Before  death,  there  was  paresis  of  the  hind  legs,  the 
appetite  had  disappeared  and  the  animal  showed  a  27  per  cent  loss 
in  weight.  The  histological  nerve  findings  were  not  so  typical  for 
experimental  beriberi,  and  controls  with  the  addition  of  vitamine  B 
were  lacking,  to  prove  that  this  condition  really  was  caused  by  lack 
of  this  substance.  Shiga  and  Kusama  (448)  have  described  a  real 
beriberi  in  monkeys,  with  anasarca,  hydropericardium  and  edema  of 
the  lungs,  while  Noe  (449)  was  unable  to  induce  beriberi  in  these 
animals  by  feeding  rice.  McCarrison  (450)  tried  then  to  produce 
beriberi  in  Macacus  sinicus,  by  feeding  autoclaved,  ground  white 
rice,  while  in  another  experiment  extra  butter  was  added.  The  first 
group  died  in  about  23.4  days  and  the  second  (with  butter),  after  15 
days.  None  of  the  animals  lived  longer  than  100  days.  Although 
typical  beriberi  or  edema  was  not  noted,  the  clinical  symptoms 
consisted  of  progressive  anemia,  gastro-intestinal  disturbances  and 
progressive  asthenia.  Stomach,  intestinal  walls  and  omentum  were 
very  thin  and  had  lost  their  fat  content.  Congested,  necrotic  and 
inflammatory  changes  were  apparent  in  the  entire  gastro-intestinal 
tract.  Aside  from  this,  there  were  degenerative  changes  in  the 
neuro-muscular  apparatus  of  the  intestines,  leading  to  the  distention 
of  the  stomach  and  to  other  parts  of  the  intestine.  Signs  of  necrosis 
were  seen  in  the  secretory  layers,  and  attenuation  of  the  elements 
protective  against  infection  was  noted.  These  findings  need  not  of 
course  be  attributed  only  to  a  lack  of  vitamine  B,  since  in  the  diet 
utilized,  vitamine  C  was  also  lacking. 

The  monkey  is  of  greater  value  in  the  study  of  scurvy,  as  was 
found  by  Hart  (451),  and  later  by  Hart  and  Lessing  (452).  Young 
monkeys  kept  on  condensed  cooked  milk,  rice  and  white  bread, 


154  THE   VITAMINES 

developed  a  condition  which  resembled  infantile  scurvy  in  its  symp- 
toms, while  in  grown  monkeys,  the  symptoms  were  more  like  those  in 
grown  people.  In  their  book,  they  gave  a  detailed  enumeration  of 
the  symptoms  and  the  pathological  findings  (with  many  illustrations), 
so  that  we  must  refer  the  reader  to  this  book  for  greater  details. 
Particularly  were  the  changes  in  the  bones  accurately  described.  The 
experiments  of  Hart  were  confirmed  by  Talbot,  Todd  and  Peterson 
(453),  and  a  few  years  later  by  Harden  and  Zilva  (454).  The  latter 
produced  scurvy  on  a  diet  containing  only  traces  of  vitamine  B,  but 
concluded  that  in  future  investigations,  it  would  seem  desirable  that 
the  diet  should  contain  sufficient  vitamine  A  and  B.  In  our  opinion, 
too,  this  is  very  important,  since  in  vitamine  studies,  it  must  be  so 
arranged  that  only  a  single  factor  is  lacking.  Harden  and  Zilva 
were  able  to  cure  the  disease  by  means  of  antiscorbutics.  In  a  later 
publication,  they  (455)  showed  that  it  took  two  months  to  develop 
scurvy  in  monkeys.  The  diet  used  by  them  consisted  of  300  grams 
of  rice,  50  grams  of  wheat  germ,  2  grams  of  salts  and  5  grams  of  butter. 
A  daily  administration  of  2  cc.  orange  juice  per  os  sufficed  to  protect 
the  animals  against  scurvy.  If  smaller  doses  were  given,  the  animals 
developed  scurvy,  with  paralysis  of  the  hind  legs,  which  appeared  to 
be  a  pseudo-paralysis.  It  is  interesting  to  note  that  the  monkey 
requires  less  vitamine  C  than  does  the  guinea  pig.  A  monkey 
weighing  from  two  to  three  kilos  needs  as  much  as  a  guinea  pig 
weighing  300  to  400  grams. 

Among  other  symptoms,  Hart  and  Lessing  (I.e.  452)  noticed  an 
exophthalmus  on  the  left  side,  with  severe  hemorrhagic  edema  of  the 
upper  eye-lids.  The  same  observation  was  also  made  by  Zilva  and 
Still  (456). 

Chemical  studies  of  the  mineral  metabolism  of  the  scorbutic 
monkey  were  made  by  Howard  and  Ingvaldsen  (457).  In  this 
case,  the  disease  was  produced  with  condensed  milk  in  4  months. 
Loss  of  salts,  usually  so  marked  in  man  and  guinea  pigs,  was  not 
noted  here. 

Harden  and  Zilva  (458)  reported  an  experiment  on  three  monkeys, 
two  years  of  age,  fed  on  250  to  300  grams  of  rice,  10  grams  of  yeast 
extract  (marmite),  2  grams  of  salts  and  4  cc.  orange  juice.  One  of 
these  animals  received  no  fat  and  no  vitamine  A,  and  developed  an 
edema  with  diarrhoea  in  289  days.  The  second  animal  received 
butter  and  lived  299  days,  whereupon  the  experiment  was  interrupted. 


THE   VITAMINES    IN   THE   ANIMAL   KINGDOM  155 

The  third  animal  was  given  olive  oil,  and  died  after  262  days  without 
characteristic  symptoms.  Butter  had  no  growth-promoting  influ- 
ence. Since  the  diet  was  very  poor  in  fat,  and  since  the  significance 
of  fat  for  monkeys  is  not  known,  the  investigators  wished  to  await 
further  results  before  saying  what  role  vitamine  A  plays  in  the 
development  of  this  edema.  However,  Hewlett  and  de  Korte  (459) 
stated  that  in  1907  he  had  described  edema  in  monkeys  which  had 
received  sufficient  milk  to  provide  for  their  vitamine  A  requirements ; 
nevertheless,  he  might  have  been  dealing  with  a  lack  of  vitamine  B. 
No  albumin  was  noted  in  the  urine  of  these  animals. 

In  conclusion,  we  wish  to  point  out  that  Chick  and  Hume  (460) 
have  described,  in  a  monkey,  a  condition  which  might  be  considered 
to  be  identical  with  human  pellagra.  The  diet  used  consisted  of 
sugar,  corn  starch,  salts  and  corn  gluten,  in  varying  amounts,  so 
that  in  this  way  more  or  less  protein  was  given.  All  of  the  three 
known  vitamines  were  present  in  the  diet.  We  shall  speak  again  of 
this  experiment  in  discussing  the  etiology  of  pellagra. 

For  the  investigators  who  are  interested  in  nutrition  experiments 
with  these  animals,  it  may  be  of  interest  to  point  out  how  monkeys 
are  raised  by  dealers  and  fed  in  our  zoological  gardens.  Schaumann 
(I.e.  2)  states  that  in  the  Institute  for  Ship  and  Tropical  Hygiene  in 
Hamburg,  monkeys  are  kept  on  a  porridge  made  of  cooked  white 
rice;  in  addition  they  receive  carob  seed,  peanuts  and  fruits.  On 
personal  inquiry  at  the  New  York  Zoological  Garden,  we  learned  that 
the  monkeys  were  given  cooked  rice,  a  cup  of  milk,  bananas,  apples, 
sweet  potatoes,  onions  and  carrots;  once  a  week  they  were  given 
some  powdered  sulphur.  The  chimpanzee  received  two  raw  eggs  in 
milk,  bananas,  two  oranges  and  a  dozen  cooked  prunes  every  two 
days.  The  monkeys  lived  for  years  on  these  diets  in  very  good 
health. 

Vitamine  requirements  of  man 

After  having  discussed  the  vitamine  and  food  requirements  of 
various  animals,  we  are  prepared  to  study  the  above  relationship  in 
man.  It  is  not  infrequently  seen  that  clinicians  are  unwilling  to 
apply  the  results  of  animal  experimentation  to  man,  and  they  are 
justified  in  being  sceptical.  Yet  we  must  emphasize  that  all  we  have 
discussed  in  the  preceding  chapters  is  based  upon  a  solid  foundation. 
We  are  dealing  here  with  the  fundamental  laws  of  cell  nutrition 


156  THE    VITAMINES 

applicable  to  all  living  organisms.  As  little  as  the  modern  clinician 
can  afford  to  ignore  the  physiological  significance  of  the  proteins  or 
of  certain  amino  acids,  just  so  little  need  he  doubt  the  importance  of 
the  vitamines  for  human  nutrition.  It  is  true  that  the  conditions 
leading  to  deficiency  diseases  in  man  are  perhaps  not  so  clear  cut  as 
in  animal  experiments,  since  in  the  latter,  we  may  choose  such  con- 
ditions as  will  permit  of  the  study  of  the  significance  of  a  single 
constituent  exactly.  In  spite  of  this,  we  do  not  yet  know  all  there 
is  to  be  known  of  the  laws  of  nutrition.  It  has  been  frequently 
necessary  to  proceed  empirically;  for  example,  we  fed  white  rice  to 
chickens  and  pigeons  without  knowing  at  the  time  that  this  food- 
stuff lacks  other  important  constituents  besides  vitamine  B.  We 
were  still  in  the  empirical  stage  when  we  wrote  our  first  edition,  but 
since  then,  vitamine  research  has  made  great  strides;  nevertheless, 
many  of  the  findings  obtained  at  that  time,  with  few  exceptions, 
still  endure  to  this  day.  Should  it  appear  later  that  the  human 
avitaminoses  are  associated  not  only  with  a  deficiency  of  the  corre- 
sponding vitamines,  but  with  other  correlated  factors,  then  the 
significance  of  vitamines  for  life  would  still  retain  its  complete 
justification.  That  the  etiology  in  human  avitaminoses  may  be  more 
complicated  than  appears  at  first  glance,  we  have  already  pointed 
out.  Should  it  happen  that  there  is  a  deficiency  of  one  vitamine, 
we  may  be  almost  certain  that  it  is  not  the  only  fault  in  the  diet 
leading  to  the  particular  disease.  Along  with  the  lack  of  one  vita- 
mine,  there  may  be  a  sub-optimal  quantity  of  other  vitamines;  there 
may  also  be  a  protein  and  salt  deficiency.  In  addition  to  this,  it  is 
possible  that  the  quantitative  relationship  of  the  individual  con- 
stituents may  not  be  correct;  it  may  likewise  be  (and  this  is  very  often 
the  case  in  practice)  that  the  vitamine  is  indeed  present  in  the  diet 
but  not  in  quite  sufficient  quantities.  Related  as  it  is  to  all  these 
factors,  we  must  not  wonder  that  the  picture  of  an  avitaminosis  often 
seems  to  us  to  be  clouded.  Taking  rickets  as  a  concrete  example, 
we  see  that  although  this  disease  is  not  of  rare  occurrence  among 
well-to-do  people,  it  nevertheless  occurs  most  frequently  in  the  poorer 
stratas  of  society,  where  poor  living  conditions  and  hereditary  factors, 
together  with  dietary  causes,  may  play  a  part.  It  is  also  not  likely 
that  the  lack  of  vitamine  A  only  is  the  cause  of  rickets,  as  is  now 
thought  to  be  the  case;  it  may  be  associated  with  a  partial 
deficiency  of  other  factors,  which  chiefly  consist  in  that  the  balance 


THE    VITAMINES    IN   THE    ANIMAL    KINGDOM 


157 


between  the  proteins  and  the  carbohydrates  is  disturbed  in  favor  of 
the  latter.  All  these  conditions  may  affect  the  clinical  picture  of 
rickets  as  we  understand  it. 

Most  of  the  pathological  conditions  which  we  designate  as  human 
deficiency  diseases  find  their  replica  in  animal  affections  already 
described.  The  diseases  have  been  produced  experimentally  and  are 
regarded  by  our  greatest  specialists  as  identical  with  human  avitam- 
inoses.  However,  this  does  not  exhaust  our  data  on  the  subject. 


IS 


JLQ         Z.S 


30         35 
DAYS 

FIG.   37.  THREE  SEPARATE  ORANGE  JUICE   (O.J.)   ADDITIONS  IN  A  PERIOD 
OF  60  DAYS  GAVE  UNIFORM  WEIGHT  INCREASES 

During  period  I,  an  increase  in  the  food  produced  no  corresponding  increase 
in  weight.  When  the  usual  quantity  of  orange  juice  (15  cc.)  was  given  at 
B,  a  second  increase  in  food  was  made.  The  effect  of  the  two  food  increases 
(dextri-maltose  D.M.)  was  distinctly  less  than  that  produced  by  the  larger 
quantity  of  orange  juice.  Orange  juice  from  which  the  B-vitamine  was 
removed  by  kaolin  showed  no  such  effect  (Byfield,  Daniels  and  Loughlin). 

Exact  experiments  have  also  been  conducted  on  man,  having  the 
convincing  value  of  a  well-planned  animal  experiment.  Thus,  we  are 
familiar  with  clinical  cases  in  which  the  food  intake  was  controlled, 
so  that  the  course  of  the  particular  disease  could  be  followed  quite 
accurately.  From  what  has  been  said,  it  is  clear  that  we  are  no  longer 
concerned  with  conjectures  in  the  classification  of  human  avitami- 
noses.  We  are  in  possession  of  proof,  which  makes  it  certain  that 
man  needs  at  least  three  vitamines — antiberiberi  (B),  antirachitic 
(A),  and  antiscorbutic  (C). 


158 


THE    VITAMINES 


Especially  from  children's  clinics,  do  we  have  observations  that 
are  in  accord  with  the  results  of  animal  experiments.  In  this  regard, 
we  wish  particularly  to  mention  the  work  of  Hess  (461),  who  studied 
the  influence  of  vitamine  C  on  the  growth  and  weight  of  infants.  To 
be  sure,  By  field,  Daniels  and  Loughlin  (I.e.  90)  believed  that  his 
results  must  have  been  due  to  the  simultaneous  presence  of  vitamine 


to 

3 


UJ 


°  4-  8  12-  16 

WEEKS 

FIG.  38.  BREAST-FED  CHILD  SHOWING  STATIONARY  WEIGHT 

Mother  received  an  addition  of  50  grams  butter  and  30  grams  turnip  juice 
(vitamines  A  and  B),  at  the  point  indicated  (Dalyell;  courtesy  of  Brit. 
Med.  J.) 

B,  but  the  value  of  the  experiment  of  Hess  is  not  lost  for  our  purpose. 
Byfield,  Daniels  and  Loughlin  were  able  to  show  that  when  children 
are  given  orange  juice  shaken  with  fuller's  earth,  thereby  removing 
vitamine  B,  there  is  no  influence  on  growth,  whereas  the  untreated 
orange  juice  has  a  far  greater  effect  than  an  addition  of  extra 
calories,  in  the  form  of  an  increased  basal  diet.  This  is  evident 
from  the  illustration  (Fig.  37).  It  is  interesting  to  note  that  this 


THE    VITAMINES    IN   THE    ANIMAL   KINGDOM 


159 


experiment  with  children  was  accompanied  by  a  similar  experiment 
with  rats,  with  identical  results.  A  second  example  of  this  kind  is 
seen  in  the  work  of  Chick  and  Dalyell  (462),  who  studied  the  effect 
of  vitamines  particularly  on  under-nourished  children  in  Vienna. 
Here,  too,  the  growth-promoting  influence  of  vitamine  C  may 


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FIG.  39.  TWINS,  BOY  AND  GIKL,  WITH  SCURVY. 

The  girl  received  at  the  point  C  an  antiscorbutic  in  the  form  of  raw  inilk; 
later  at  AC,  lemon  or  turnip  juice,  with  butter  or  cod  liver  oil  were  given. 
The  boy  was  suffering  from  subacute  scurvy  and  did  not  get  any  antiscorbutic 
except  at  the  place  marked  C,  when  his  growth  markedly  improved;  he  suffered 
also  from  rickets.  (Dalyell;  courtesy  of  Brit.  Med.  J.) 

perhaps  be  explained  by  the  apparent  presence  therein  of  vitamine 
B.  The  administration  of  vitamine  to  the  nursing  mother,  with 
effect  on  the  growth  of  the  child,  can  find  its  parallel  only  in  the 
most  exact  animal  experiment. 

Logically,  we  should  begin  here  with  the  description  of  human 
avitaminoses,  like  beriberi,  scurvy,  rickets,  osteomalacia,  and  some 


160  THE    VITAMINES 

which  are  not  yet  classified  as  such,  like  pellagra  and  war  edema. 
Other  conditions  also,  like  ophthalmia,  dental  caries,  and  some 
infectious  and  constitutional  conditions,  the  progress  of  which  seems 
to  be  influenced  by  vitamine  additions,  should  be  mentioned  here. 
The  application  to  human  pathology  is  obviously  the  objective  point 
of  the  entire  vitamine  problem.  We  believe,  therefore,  that  after  the 
demonstration  of  the  chemistry  and  physiology  of  the  vitamines,  as 
well  as  the  vitamine  content  of  the  various  foodstuffs,  and  the  influ- 
ence of  the  treatment  these  foodstuffs  undergo  before  consumption, 
the  reader  will  be  better  able  to  perceive  the  real  significance  of  the 
vitamines  in  human  pathology. 


PART  II 

Chemistry,  Physiology  and  Pharmacology 
of  the  Vitamines 


CHAPTER  I 
THE  ANTIBERIBERI  VITAMINE — VITAMINE  B 

In  the  foregoing  chapters,  we  have  discussed  facts  which  permitted 
of  the  assumption  of  the  existence  of  at  least  three  different  vitamine 
types.  A  great  number  of  investigations  of  a  chemical  nature  have 
already  made  their  appearance,  dealing  with  the  characteristics  of 
these  important  substances;  it  may  indeed  be  said  that  no  single 
division  of  physiological  chemistry  has  latterly  been  productive  of 
more  publications  than  that  of  the  vitamines.  The  reason  is  that 
new  facts  and  observations  are  obtained  with  an  ease  that  is  truly 
astounding.  Because  of  this,  many  investigators  have  failed  to 
penetrate  more  deeply  into  this  subject.  They  have  satisfied  them- 
selves, as  a  rule,  with  results  obtained  by  indirect  conclusions.  This 
condition  of  marking  time  and  useless  quibbling  provides  an  excellent 
opportunity  for  the  representatives  of  destructive  criticism  which, 
here  more  than  in  any  other  field  of  research,  is  effective  in  hindering 
the  solution  of  our  problem.  In  spite  of  these  difficulties,  the  knowl- 
edge of  the  nature  of  the  vitamines  progresses  gradually,  and  we  may 
hope  that  with  the  resumption  of  scientific  research,  after  the  war, 
the  desired  goal — the  chemical  isolation  of  the  vitamines — is  not  far 
distant. 

A  question  which  we  must  touch  upon  is  the  comparison  often 
made  between  the  vitamines  and  the  ferments,  a  relationship 
specially  emphasized  by  Seidell  (462a).  Since  the  nature  of  the 
ferments,  in  spite  of  the  many  years  of  study,  still  remains  to  be 
cleared  up,  this  comparison  raises  a  poor  prognosis  as  to  the  outcome 
of  our  vitamine  investigations.  However,  we  find  many  far  reaching 
differences  between  these  two  classes  of  substances.  Whereas  we 
may  observe  in  ferments  that  in  a  temperature  interval  between  50° 
and  70°  C.,  there  is  a  sudden  snapping,  like  the  over-wound  main 
spring  in  a  clock,  we  note  in  vitamines  that  the  inactivation  proceeds 
gradually  with  an  increase  in  temperature.  In  general,  the  complete 
destruction  of  vitamines  is  seen  after  a  much  higher  temperature 
than  in  ferments.  The  vitamines  appear  to  us,  as  compared  with 
ferments,  to  have  a  relatively  simple  structure,  and  it  would  be  highly 
desirable  to  have  more  pure  organic  chemists  take  up  this  problem. 

163 


164  THE   VITAMINES 

We  come  now  to  the  discussion  of  the  difficulties  involved  in  the 
field  of  vitamine  fractionation.  Starting  from  any  crude  material 
and  subjecting  it  to  a  variety  of  chemical  and  physical  manipulations, 
we  usually  find  that  the  vitamines  are  quite  resistant.  If  now  a 
chemical  fractionation  is  undertaken,  we  note  that  some  procedures 
cause  either  a  partial  or  total  disappearance  of  vitamine  activity. 
What  then  is  there  in  the  natural  combination  which  protects  the 
vitamines  against  destruction?  It  has  long  been  thought  that  it 
was  an  acid  which  served  to  protect  these  substances,  but  this  is 
apparently  only  partially  the  case.  A  second  possibility  is  that  the 
natural  combination  is  protective  against  oxidation;  this  may  be 
tested  experimentally  and  will  be  spoken  of  later. 

The  main  difficulty  of  isolation,  in  our  opinion,  is  that  in  most  of 
the  starting  material  used  till  now,  the  vitamines  are  associated  with 
large  amounts  of  inactive  substances.  When  we  mix  a  small  quantity 
of  a  known  substance  with  a  large  quantity  of  another  known  sub- 
tance,  we  find  it  hard  to  effect  a  separation.  Here,  too,  we  are 
dealing  with  the  law  of  mass  action,  and  only  those  substances  which 
occur  in  large  amounts  in  the  mixture  may  be  easily  separated.  Now, 
if  we  recall  that  we  are  dealing  with  cellular  constituents,  the  chemical 
nature  of  which  is  still  insufficiently  known,  then  it  is  clear  that  our 
problem  becomes  much  more  complicated.  Just  how  little  the  con- 
stituents of  the  cells  are  known  becomes  more  evident  when  we 
attempt  a  fractionation  of  the  vitamines;  we  immediately  encounter 
cell  components  totally  unknown  to  us.  It  is  sought  to  avoid  these 
difficulties  by  the  choice  of  starting  material  which,  in  addition  to  a 
definite  vitamine  content,  has  less  inactive  substances.  Even  this 
procedure  is  uncertain,  since  upon  concentration  of  the  vitamines, 
there  is  a  corresponding  concentration  of  the  impurities.  There  is 
no  method  whereby  we  may,  in  one  operation,  remove  the  vitamines 
from  the  bulk  of  its  associated  impurities,  and  which  would  be 
applicable  to  a  large  quantity  of  crude  material.  Besides,  it  is  not 
impossible  that  the  delay  attending  the  usul  chemical  procedures 
may  have  some  destructive  influence  on  the  vitamines.  For  example, 
if  it  should  be  necessary  to  exclude  air,  it  is  not  very  difficult  to 
provide  for  this  for  a  few  days,  but  it  is  another  matter  when  the 
procedures  employed  involve  weeks  instead  of  days.  It  is  necessary, 
therefore,  that  we  should  try  to  develop  some  color  reaction  or 
pharmacological  test  for  the  demonstration  of  the  vitamines,  which 


THE   ANTIBERIBEKI   VITAMINE  165 

may  provide  an  answer  for  the  question  as  to  which  fraction  contains 
the  vitamine.  It  sometimes  takes  weeks  before  we  are  certain  of 
this,  and  during  this  time  the  fractions  obtained  must  await  further 
manipulations.  The  means  used  till  now  for  the  study  of  the  nature 
of  the  vitamines  (almost  exclusively  the  more  stable  vitamine  B) 
are  as  follows: 

1.  Fractionation  of  the  active  material  according  to  the  known 
methods. 

2.  Attempts  to  separate  the  active  material  by  solubility   and 
selective  adsorption. 

3.  Testing  of  vitamine-like  action  of  combinations  of  known  or 
unknown  composition. 

4.  Synthetic  experiments  with  substances  obtained  in  vitamine 
work. 

Up  to  the  present,  none  of  these  four  procedures  has  yielded  very 
substantial  results,  although  the  first  of  these  methods,  which  is 
most  direct,  has  been  productive  of  the  most  instructive  findings. 
By  this  means,  some  precipitation  reactions  have  been  found  which 
lead  to  a  vitamine  fraction  which  appears  to  be  quite  simple  in  com- 
parison with  the  complexity  of  the  starting  material.  From  these 
fractions,  there  have  been  isolated  well  defined  substances  which 
possess  a  very  marked  curative  action  while  in  the  impure  state,  but 
which,  upon  further  purification,  lose  much  if  not  all  of  their  activity. 
Nevertheless,  it  has  been  possible  to  isolate  a  substance  that  has  been 
crystallized  to  constant  melting  point,  at  the  same  time  retaining  a  good 
deal  of  its  activity;  this  was  the  case  with  our  work  on  yeast.  How- 
ever, the  objection  is  justified  that  the  vitamine  is  present  in 
such  a  small  quantity  that  it  has  no  effect  on  the  melting  point.  We 
must  determine  now  whether  we  are  dealing  with  inactive  material 
that  has  adsorbed  the  vitamine,  or  with  one  or  more  cleavage  prod- 
ucts serving  as  the  inactive  basic  structure  for  the  vitamines.  It  is 
obviously  very  unlikely  that  the  vitamines  should  disappear  without 
leaving  a  trace.  When  radium,  for  example,  undergoes  complete 
decomposition,  the  end  product  is  inactive  lead.  It  is  not  impossible 
that  in  vitamines  we  encounter  a  similar  problem.  Our  view  is 
further  strengthened  in  this  respect  when  amongst  the  substances 
usually  accompanying  vitamine  B,  we  constantly  meet  with  products, 
having  the  pyridine  ring,  which  has  been  recognized  as  a  cell  con- 
stituent for  the  first  time  in  vitamine  work.  If  we  were  certain 


166  THE   VITAMINES 

that  these  pyridine  derivatives  are  indicative  of  decomposition 
products  of  vitamines,  then  it  would  be  possible  to  demonstrate  their 
formula  and  chemical  constitution  with  comparative  ease;  subse- 
quently the  synthesis  could  be  made.  Although  the  amount  of  these 
pyridine  derivatives  is  small  enough  to  permit  of  a  possible  relation- 
ship with  the  vitamines,  our  facts  are  not  sufficiently  certain  to 
proceed  to  such  work,  since  in  the  end  we  may  be  dealing  with 
incidental  impurities. 

The  second  method  of  isolation,  by  extraction  with  various  solvents 
or  treatment  with  adsorptive  agents,  has  given  some  interesting 
results,  though  it  could  finally  be  shown  that  such  fractions  still 
contained  complexes.  Hence  a  further  fractionation  would  again  be 
necessary,  whereby  the  above  mentioned  difficulties  would  again  have 
to  be  dealt  with. 

The  third  way,  naturally  only  adapted  to  exclude  from  our  con- 
sideration substances  of  known  composition,  did  not  prove  of  value 
in  the  studies  of  the  nature  of  vitamines.  Obviously,  a  positive 
result  might  have  been  due  to  a  contamination  with  vitamine,  or  to 
a  secondary  effect  that  might  be  able  to  simulate  a  vitamine  action. 

The  fourth,  or  synthetic,  procedure  has  yielded  no  results  till 
now,  since  the  investigations  have  been  based  on  weak  foundations. 
What  then  is  the  best  method  for  the  chemical  study  of  the  vita- 
mines?  First  of  all,  we  must  stabilize  the  vitamines,  steps  towards 
which  have  already  been  taken.  If  this  should  prove  to  be  impracti- 
cable, we  must  look  for  another  vitamine  in  Nature  which  is  more 
stable.  We  shall  see  directly  that  in  general,  vitamine  B  is  more 
stable  than  the  two  other  vitamines.  It  would  suffice  if,  after  the 
chemical  purification  of  this  substance,  there  remained  a  single 
function  which  would  characterize  the  substance  as  a  vitamine.  We 
have  already  learned  that  all  three  vitamine  types  are  present  in 
seeds,  or  that  they  may  eventually  arise  through  chemical  re-arrange- 
ment. Although  we  have  no  positive  proof  that  the  three  vitamine 
types  are  related  chemically,  it  is  not  impossible  that  the  identifica- 
tion of  a  single  vitamine  may  be  of  immeasurable  help  in  the  study 
of  the  other  vitamine  types. 

The  vitamines  have  this  in  common,  that  they  are  all  necessary 
to  complete  a  synthetic  diet.  Further,  none  of  the  vitamines  can  be 
identified  with  any  of  the  dietary  constituents  already  known;  they 
are  active  in  very  small  quantities  and  none  of  them  possess  any  too 


THE   ANTIBERIBERI   VITAMINE  167 

great  stability.  It  is  possible,  on  the  one  hand,  that  we  may  need 
new  chemical  methods  in  order  to  discover  their  chemical  nature; 
on  the  other  hand,  it  is  not  unlikely  that  our  older  methods,  which 
have  been  of  such  use  to  us  in  the  identification  and  isolation  of  so 
many  naturally  occurring  products,  may  lead  us  to  the  desired  goal. 
We  have  already  pointed  out  the  status  of  this  question,  up  to 
1911,  in  the  historical  part.  We  stated  that  till  then,  the  investiga- 
tors in  this  field  were  in  doubt  as  to  whether  the  factor  curing  beriberi 
was  really  a  chemical  substance.  We  have  shown,  together  with 
Cooper  (I.e.  68)  that  when  pressed  yeast  is  hydrolyzed  by  boiling 
vigorously  with  20  per  cent  sulphuric  acid  for  24  hours,  a  id  then  the 
sulphuric  acid  completely  removed  with  baryta,  the  evaporated 
filtrate  still  exhibited  a  very  marked  curative  action.  This  stability 
in  the  presence  of  acid  makes  it  appear  that  the  substance  in  question 
is  an  organic  base,  which  would  distinguish  itself  by  the  known 
chemical  characteristics  of  this  class  of  substances.  Based  upon  this 
assumption,  we  (463)  undertook  a  systematic  investigation  of  rice 
polishings. 

CHEMICAL    INVESTIGATION   OF    RICE    POLISHINGS 

Since  no  chemical  reaction  was  known  for  the  demonstration  of 
this  vitamine,  every  one  of  the  investigated  fractions  was  tested  on 
beriberi  pigeons.  The  stage  at  which  vitamine  was  administered 
(mostly  given  per  os  in  the  beginning)  was  the  appearance  of  con- 
tractions of  the  neck,  wings  and  legs.  Left  to  themselves,  the 
pigeons  died  in  about  12  hours  from  the  onset  of  the  above  symptoms. 

A  series  of  preliminary  experiments  were  made  in  order  to  secure 
an  active  solution  of  the  simplest  possible  composition.  This  was 
obtained  by  shaking  rice  polishings,  which  consist  chiefly  of  cellulose, 
phytin  and  fat,  with  alcohol  saturated  up  to  a  certain  point  with 
gaseous  hydrochloric  acid.  This  procedure  possessed  the  advantage 
that  the  solution  was  completely  freed  of  alcohol  insoluble  material. 
The  alcoholic  solution  was  then  concentrated  in  vacuum,  yielding  a 
fatty  residue.  This  residue  was  melted  on  the  water  bath,  extracted 
with  hot  water,  and  the  layers  separated  in  a  separatory  funnel 
while  still  hot.  The  watery  extract,  which  was  very  active,  was 
treated  with  sulphuric  acid  till  a  5  per  cent  solution  was  obtained, 
and  then  completely  precipitated  with  a  50  per  cent  phosphotungstic 
acid  solution.  The  precipitate  was  decomposed  with  baryta  and  the 


168  THE    VITAMINES 

resulting  filtrate,  after  removal  of  the  excess  baryta,  was  tested  for 
its  curative  power  on  beriberi  pigeons.  The  solution  was  very  active 
and  was  free  of  phosphorus,  protein,  and  carbohydrates  (I.e.  463). 
The  first  difficulty  encountered  was  that  of  determining  the  exact 
dose  for  a  sick  pigeon.  As  will  be  shown  later,  the  solution  con- 
tained much  free  choline  which  is  very  toxic  for  pigeons.  This 
difficulty  could  be  overcome  by  calculating  the  dosage  on  the  basis  of 
the  quantity  of  rice  polishings  used.  With  every  additional  fraction, 
the  naturally  occurring  loss  must  be  made  up  by  an  increased  dosage. 

The  fact  that  the  solution  was  phosphorus-free  served  as  a  final 
blow  to  Schaumann's  hypothesis  regarding  the  lack  of  phosphorus, 
since  it  was  demonstrated  for  the  first  time  that  a  solution  completely 
free  from  phosphorus,  was  active.  Since  the  phosphotungstic  acid 
filtrate  was  absolutely  inactive,  all  of  the  active  substance  must  have 
been  contained  in  the  precipitate. 

The  usual  methods  were  used  in  the  working  up  of  such  precipitates. 
The  resultant  solution,  which  contained  a  large  amount  of  potassium 
and  hence  reacted  strongly  alkaline,  was  neutralized  with  hydro- 
chloric acid  and  then  concentrated  to  a  syrup  in  vacuum.  The 
residue  was  taken  up  in  alcohol,  leaving  the  greatest  part  of  the 
potassium  chloride  behind.  The  alcoholic  solution  was  precipitated 
by  sublimate,  the  precipitate  being  recrystallized  from  water,  with 
the  addition  of  some  sublimate.  The  crystals  were  then  suspended 
in  water  and  decomposed  with  hydrogen  sulfide.  This  solution,  as 
well  as  that  obtained  after  the  elimination  of  excessive  sublimate,  was 
active  (the  filtrate,  however,  being  more  so).  Hence  it  was  not 
possible  to  effect  a  separation  with  sublimate.  The  solution  obtained 
from  the  precipitate  was  concentrated  in  vacuum,  the  residue  dis- 
solved in  alcohol  and  precipitated  with  an  alcoholic  solution  of 
platinic  chloride.  A  series  of  fractions  were  obtained  which,  on 
analysis,  proved  to  be  pure  choline.  The  alcoholic  solution  was  still 
active  after  complete  elimination  of  choline.  It  could  not  be  doubted 
therefore  that  the  curative  substance  did  not  belong  to  the  choline 
group.  In  the  mother  liquor  remaining  after  precipitation  with 
sublimate,  various  precipitants  were  tried  in  order  to  precipitate  the 
active  substance. 

For  this  purpose,  silver  nitrate  in  alkaline  solution  was  used  by  the 
author  (corresponding  to  the  precipitant  for  the  histidine  group). 
The  solution  was  freed  from  chlorine  by  silver  sulfate.  After  the 


THE   ANTIBERIBERI   VITAMINE  169 

removal  of  silver  and  sulphuric  acid,  the  solution  was  acidified  with 
nitric  acid,  and  then  treated  with  silver  nitrate  till  it  gave  a  brown 
precipitate  with  baryta. 

A  saturated  baryta  solution  was  then  added  so  long  as  a  drop  of 
the  clear  solution  gave  even  a  slight  white  precipitate  on  addition  of 
ammoniacal  silver  nitrate  solution.  The  precipitate  was  then 
washed  free  of  nitric  acid,  decomposed  with  hydrogen  sulphide,  the 
last  traces  of  baryta  removed  with  very  dilute  sulphuric  acid,  and 
concentrated  in  vacuum.  The  solution  thus  obtained  was  very 
active.  By  means  of  the  gradual  evaporation  of  the  solution  in  the 
desiccator,  with  the  addition  of  alcohol,  a  colorless  crystalline  sub- 
stance was  obtained.  This  weighed  0.4  gram  out  of  50  kilos  of  rice 
polishings  (secured  from  Malay).  The  very  small  yield  prevented 
further  work,  it  being  possible  only  to  make  a  series  of  reactions. 
The  needle-like  crystals,  melted  sharply  at  233°C.  (uncorrected)  and 
were  rather  insoluble  in  cold  water  and  cold  alcohol.  At  that  time, 
no  other  substance  could  be  found  in  this  fraction. 

As  to  the  therapeutic  action  of  the  product,  we  expressed  ourselves 
only  with  great  caution,  but  beyond  all  doubt  there  was  a  curative 
action  present.  The  amount  used  for  a  curative  dose  contained  only 
four  milligrams  of  nitrogen.  Smaller  doses  were  not  tried  at  the 
time. 

In  a  later  publication  (I.e.  62),  we  called  this  active  substance 
"beriberi  vitamine,"  to  make  it  evident  that  it  contained  nitrogen 
and  was  indispensible  for  life. 

To  illustrate  our  procedure,  we  append  a  schematic  diagram. 

Rice  polishings >    Alcohol  extract >    Watery  extract 


Phosphotungstic  acid  precipitate    >    Filtrate 


/     \ 


KC1          Sublimate         Silver  nitrate  and  baryta. 

I  I 

Choline  Vitamine. 

Still  later,  we  attempted  to  simplify  our  procedure  (I.e.  324).  In 
the  first  place,  the  rice  polishings  were  extracted  with  ordinary  alcohol 
instead  of  acid  alcohol.  The  watery  extract  was  then  immediately 


170  THE   VITAMINES 

precipitated  with  silver  and  baryta,  instead  of  phosphotungstic  acid. 
We  observed  that  this  extract  behaved  quite  differently  from  the 
previous  one;  it  appeared  to  contain  less  potassium  and  no  free 
choline.  On  decomposing  the  silver  precipitate,  a  substance  was 
obtained  (not  the  vitamine  fraction),  which  melted  at  231°C.  and 
was  identified  as  allantoine.  Schaumann  (464)  described  the  isola- 
tion of  a  substance  containing  nitrogen,  which  was  perhaps  allantoine 
but  was  not  investigated  either  chemically  or  therapeutically. 

We  have  therefore  isolated  from  rice  polishings  two  known 
nitrogenous  substances,  allantoine  and  choline,  and  the  vitamine 
fraction.  The  method  described  in  our  first  paper  on  rice  polishings 
seems  to  be  peculiarly  adapted  for  the  isolation  of  the  vitamine 
fraction,  although  it  has  shown  the  problem  to  be  more  complicated 
than  was  apparent  at  first  sight.  We  met  with  particular  difficulty 
when  the  rice  polishings  were  not  extracted  in  the  laboratory  but  in 
a  chemical  factory,  and  in  large  quantities.  The  investigation  was 
begun  with  380  kilos  of  rice  polishings  (465)  using  the  method 
described  in  our  earlier  work  on  the  same  material.  For  technical 
reasons,  however,  it  seemed  necessary  to  remove  the  fat  first.  Ordi- 
nary instead  of  hydrochloric  acid  alcohol  was  used,  and  the  evaporated 
extract  was  hydrolyzed  for  two  hours  with  5  per  cent  sulphuric  acid. 
In  other  respects  the  procedure  was  the  same  as  above;  the  individual 
fractions  were  not  investigated  but  careful  experiments  were  made 
with  the  vitamine  fraction.  The  other  substances,  of  no  direct 
concern  in  our  problem,  were  kept  for  later  work.  We  thought  at 
first  that  we  had  isolated  two  different  substances  by  means  of  frac- 
tional crystallization  of  the  vitamine  fraction,  although  they  seemed 
to  crystallize  out  together,  especially  when  the  concentration  was  too 
great.  When  the  first  fraction  was  recrystallized,  needle-like  crystals 
were  obtained  which  melted  at  233°C.  (uncorrected)  and  gave  the 
following  figures  on  combustion  according  to  the  method  of  Pregl: 

4.796  mgm.  substance  yielded  10.34  mgm.  CO2  and  1.685  mgm.  H2O  corre- 
sponding to  58.80  per  cent  C  and  3.93  per  cent  H. 

4.212  mgm.  substance  yielded  9.095  mgm.  CO2  and  1.465  mgm.  H2O, corre- 
sponding to  58.89  per  cent  C  and  3.89  per  cent  H. 

4.367  mgm.  yielded  (at  713  mm.  18°C.)  0.420  cc.  N  corresponding  to  10.58  per 
cent  N. 

Only  a  small  part  of  the. nitrogen  could  be  determined  by  Van 
Slyke's  method.  Nitrogen  could  not  be  estimated  by  KjeldahPs 


THE   ANTIBERIBERI   VITAMINE  171 

method.  From  the  above  figures,  the  following  formula  was  calcu- 
lated: C26H2oN409.  Still,  as  we  shall  see  later,  these  figures  are 
almost  equally  applicable  for  the  formula  of  nicotinic  acid,  C6H5O2N, 
containing  58.3  per  cent  C,  4.08  per  cent  H,  and  11.34  per  cent  N. 
The  substance  seems  to  have  been  identical  with  that  found  in  this 
fraction  earlier  (I.e.  463).  While  fche  weight  of  the  original  crude 
product  was  2.5  grams,  that  of  the  recrystallized  substance  was  1.8 
grams.  On  additional  purification,  the  melting  point  was  no  higher. 
The  mother  liquor  was  further  fractionated,  yielding  a  substance 
which  appeared  to  be  more  soluble  in  water  than  the  first.  After 
many  recrystallizations,  it  melted  at  234°C.  (uncorrected) .  When 
treated  with  a  watery  solution  of  picric  acid,  this  substance  yielded 
a  somewhat  insoluble  pi  crate,  which  could  be  recrystallized  from 
water,  melted  at  218°C.  (uncorrected),  was  light  yellow  in  color  and 
crystallized  in '  prisms  and  plates.  The  following  results  were 
obtained  on  analysis: 

4.217  mgm.  yielded  9.625  mgm.  CO2  and  1.48  mgm.  H2O;  58.37  per  cent  C,  3.93 

per  cent  H. 
4.276  mgm.  yielded  9.16  mgm.  CO2  and  1.55  mgm.  H2O;  58.45  per   cent   C, 

4.06  per  cent  H. 

3.11    mgm.  yielded  0.315  cc.  N  (714  mm.  19.5°C.);  11.11  per  cent  N. 
3.608  mgm.  yielded  0.362  cc.  N  (712  mm.  19.5°C.);  10.97  per  cent  N. 

For  the  formula  C6H502N  (123.05)  there  is  58.3  per  cent  C,  4.08 
per  cent  H,  and  11.34  per  cent  N.  The  picrate  had  the  following 
composition : 

4.471  mgm.  yielded  6.74  mgm.  CO2  and  0.895  mgm.  H2O,  41.12  per  cent  C; 

2.24  per  cent  H. 
2.404  mgm.  yielded  0.349  cc.  N  (707  mm.  19°C.);  15.79  per  cent  N. 

These  figures  correspond  to  the  formula  Ci2H809N4  (352.12), 
which  contains  40.9  per  cent  C,  2.29  per  cent  H,  and  15.91  per  cent  N. 
The  composition  of  the  substance,  as  well  as  its  picrate,  characterizes 
it  as  nicotinic  acid  (m-pyridine  carbonic  acid).  This  substance  was 
first  found  in  Nature  by  Suzuki,  Shimamura  and  Odake  (466),  and 
was  later  described  in  detail  by  Suzuki  and  Matsunaga  (467).  We 
shall  see  later  that  the  same  substance  was  found  by  us  in  the  vita- 
mine  fraction  of  yeast. 

From  the  mother  liquor  of  the  nicotinic  acid,  another  substance 
was  isolated  which  gave  the  Millon  reaction  in  the  crude  state. 


172  THE   VITAMINES 

Because  of  the  small  yield,  it  could  not  be  investigated  further.  After 
the  publication  of  our  findings,  Schaumann  (468)  undertook  some 
experiments  with  rice  polishings,  using  our  method.  He  obtained 
a  small  amount  of  a  crystalline  substance  from  the  sublimate  fraction, 
and  demonstrated  that  it  was  active.  He  was  able  to  confirm  our 
work,  but  he  could  not  effect  a  complete  isolation.  Later,  a  great 
number  of  investigators  tried  to  verify  our  findings  and  succeeded, 
at  least  as  far  as  the  vitamine  fraction  was  concerned.  Among  these, 
we  wish  to  mention  Wellman,  Eustis  and  Scott  (469),  as  well  as 
Eustis  and  Scott  (470).  Vedder  and  R.  R.  Williams  (471)  obtained, 
with  our  method,  a  crystalline  base  that  was  therapeutically  active. 
They  found  that  unhydrolyzed  extracts  acted  more  slowly  than  the 
hydrolyzed.  Besides  this,  they  observed  that  the  vitamine  was  not 
completely  soluble  in  95  per  cent  alcohol,  and  was  destroyed  by  strong 
alkali;  it  appeared  also  that  it  could  not  be  precipitated  by  basic 
lead  acetate.  The  same  results  were  obtained  by  R.  R.  Williams  and 
Saleeby  (472),  substantiating  the  silver  nitrate-baryta  method. 
Later,  Williams  (473)  described  various  modifications  of  our  method. 
Issoglio  (474)  also  found  that  phosphotungstic  acid  precipitates  the 
vitamine.  Subsequently,  the  isolation  of  vitamine  from  rice  polish- 
ings  was  attempted  by  Kondo  and  Gomi  (475)  and  by  Murai  (476). * 
Brill  (479)  tried  to  concentrate  the  vitamine  by  adsorption  with 
infusorial  earth,  but  this  procedure  was  not  very  suitable.  Fraser 
and  Stanton  (480)  also  conducted  some  experiments,  attempting  to 
extract  and  isolate  the  vitamine. 

A  much  more  interesting  paper  was  published  in  1912  by  Suzuki, 
Shimamura  and  Odake  (I.e.  466).  They  extracted  fat-free  rice 

1  Some  patents  have  been  taken  out  on  the  isolation  of  vitamines  from  rice 
polishings.  That  of  Tsuzuki  (477)  was  nothing  more  than  a  concentrated 
alcoholic  extract  of  rice  polishings.  In  addition,  there  is  the  patent  of  Gams 
and  Schreiber  (478)  and  the  identical  patent  of  the  Society  for  Industrial 
Chemistry  at  Basle,  which  briefly  is  as  follows:  the  purification  of  the  extract 
of  rice  polishings,  which  is  supposed  to  contain  very  little  inactive  material, 
consisted  of  a  precipitation  with  lead  acetate,  first  in  acid  and  then  in  neutral 
solution.  Then  the  filtrate  was  precipitated  by  alkaloidal  reagents,  like 
phosphotungstic  acid  or  oxalic  acid,  and  the  resulting  precipitate  decom- 
posed in  a  watery  solution  with  calcium  carbonate.  By  this  means,  a  syrup 
was  obtained  which  was  soluble  in  methyl  but  not  in  absolute  ethyl  alcohol, 
ether  and  acetone;  it  gave  a  strong  diazo  reaction.  The  product  is  sold  under 
the  name  of  "Orypan." 


THE  ANTIBERIBERI  VITAMINE  173 

polishings  with  alcohol  in  a  reflux  condenser  for  three  hours,  using 
fresh  solvent,  till  the  extraction  was  complete.  The  alcoholic 
extracts  were  combined  and  concentrated.  The  residue  was  diluted 
with  water,  sulphuric  acid  added  (till  a  3  per  cent  solution  resulted) 
and  precipitated  with  30  per  cent  phosphotungstic  acid.  The  solution 
obtained  by  decomposing  the  phosphotungstic  acid  precipitate  was 
very  active  and  was  called  " crude  oryzanin  I."  The  yield  was  1.2 
grams  from  300  grams  of  fat-free  rice  polishings.  This  fraction  cured 
pigeons  promptly  in  amounts  of  3  to  4  centigrams,  while  the  filtrate 
of  the  phosphotungstic  acid  precipitate  was  entirely  inactive.  The 
active  fraction  was  soluble  in  water  and  alcohol,  was  strongly  acid 
and  gave  a  marked  Millon  and  diazo  reaction.  On  the  addition  of 
lead  acetate,  a  slight  precipitate  was  obtained,  increasing  on  the 
addition  of  ammonia.  The  substance  was  partially  precipitated 
by  sublimate,  mercuric  acetate,  and  mercuric  nitrate.  An  attempt 
to  separate  this  active  fraction  resulted  in  a  complete  loss  of 
activity.  This  was  never  the  case  in  our  experiments;  the  vitamine 
was  very  resistant  to  acids. 

The  hydrolysis  was  accomplished  by  heating  for  two  hours  with 
3  per  cent  hydrochloric  acid.  On  cooling,  yellowish  brown  crystals 
separated  out,  which  were  re  crystallized  from  hot  alcohol.  In 
this  way  two  substances  were  obtained,  the  first  of  which  was  less 
soluble  than  the  other.  Both  were  difficultly  soluble  in  water  and 
gave  an  acid  reaction;  they  wrere  soluble  in  alkali,  and  could  be 
reprecipitated  on  the  addition  of  acid.  On  analysis,  the  first  product 
gave  the  formula  CisHie^Og  and  was  called  a-acid;  the  second  prod- 
uct gave  the  formula  CioH8N04  and  was  called  0-acid;  both  showed 
the  Millon  and  diazo  reactions.  Unfortunately,  the  melting  point  and 
the  investigation  of  the  derivatives  was  not  recorded  in  the  report 
of  the  Japanese  investigators.  From  the  hydrolyzate,  choline  and 
nicotinic  acid  (m.p.214°C.)  could  be  isolated  as  picrates;  glucose 
was  also  found.  One  hundred  parts  of  crude  oryzanin  gave  10  parts 
of  a-  and  /3-acids,  30  parts  of  choline  and  nicotinic  acid,  and  23 
parts  of  glucose.  One  gram  of  crude  oryzanin  yielded  0.044  gram 
nitrogen,  of  which  0.035  gram  could  be  precipitated  by  phospho- 
tungstic acid,  of  0.009  gram  was  in  the  form  of  a-  and  /3-acids. 

As  this  composition  shows,  the  investigation  of  rice  polishings  by 
the  Japanese  workers  yielded  fundamentally  different  results  from 
ours.  The  crude  oryzanin  I.  was  then  subjected  to  further 


174  THE   VITAMINES 

purification;  it  was  dissolved  in  water  and  precipitated  with  a 
20  per  cent  tannin  solution  till  only  a  slight  cloudiness  was  seen. 
The  precipitate  was  filtered  off  and  quickly  washed  with  a  1 
per  cent  tannin  solution.  It  was  then  rubbed  up  in  a  mortar 
with  3  per  cent  sulphuric  acid  till  a  complete  solution  resulted. 
This  was  treated  with  an  excess  of  baryta,  the  precipitate  filtered 
off,  and  the  excess  of  baryta  removed  from  the  filtrate  with 
dilute  sulphuric  acid.  The  solution  was  shaken  out  with  ether  and 
concentrated.  In  this  way,  a  preparation  was  obtained,  called  crude 
oryzanin  II.,  which  was  three  times  as  active  as  the  first  preparation. 
When  a  concentrated  watery  solution  of  this  preparation  was 
rubbed  up  with  a  slight  excess  of  dilute  picric  acid,  a  flocculent 
precipitate  settled  out  which  became  crystalline  on  standing  in  the 
cold.  These  crystals  still  occluded  some  nicotinic  acid  picrate,  but 
with  careful  technique  the  latter  remained  in  solution.  The  oryzanin 
picrate  was  recrystallized  by  dissolving  in  cold  acetone  and  allowing 
this  to  evaporate  in  the  dessicator;  yellowish  brown  microscopic 
needles,  grouped  in  star  formation,  were  obtained.  An  amount 
corresponding  to  two  centigrams  of  picrate  was  very  active  for 
pigeons;  the  substance  was  given  only  to  two  pigeons.  The  amount 
of  picrate  obtained  was  so  slight  that  there  was  not  enough  for  a 
melting  point.  The  question  as  to  whether  the  pure  oryzanin  would 
give  the  same  decomposition  products  as  oryzanin  I.  was  therefore 
left  open  by  the  authors.  Since  the  publication  of  this  work  in 
1912,  nothing  of  a  corroborative  nature  has  been  printed  by  the 
Japanese  investigators  (at  least  to  our  knowledge).  In  the  mean- 
time, Drummond  and  Funk  (481)  tried  to  confirm  the  above  findings, 
but  all  attempts  to  isolate  the  substance  as  a  picrate  failed.  Above 
all,  it  was  evident  that  an  extract  of  rice  polishings  is  still  quite  a 
complicated  mixture.  Of  the  many  substances  it  contains,  we  were 
able  to  isolate,  besides  the  previously  mentioned  choline,  allantoine 
and  nicotinic  acid,  also  betaine,  adenine,  guanine  and  apparently 
guanidine.  An  observation,  made  accidently,  showed  us  how  careful 
one  must  be  in  drawing  conclusions  from  such  fractionations.  In  this 
instance,  an  apparently  pure  substance  was  isolated,  which  had  a 
constant  melting  point  on  recrystallization.  The  substance  consisted 
of  betaine  and  nicotinic  acid,  which  could  not  be  separated  from  each 
other  by  recrystallization.  Only  when  the  nicotinic  acid  was 
separated  as  a  copper  salt,  was  the  betaine  apparent.  It  was  also 


THE   ANTIBERIBERI   VITAMINS  175 

clear  from  this  work,  that  Barger  (482)  was  correct  in  his  statement 
that  the  substance  described  by  us  in  1913,  to  which  we  then  gave  the 
formula  C26H2oN4O9,  was  in  reality  nicotinic  acid.  However,  the 
view  expressed  in  the  Report  of  the  Medical  Research  Committee 
(I.e.  333),  that  the  substance  isolated  was  nicotinic  acid  contami- 
nated with  vitamine  is  erroneous,  since  the  analysis  indicated,  at  the 
time,  pure  nicotinic  acid  for  which  no  curative  action  was  claimed. 
Summarizing  our  work  with  rice  polishings,  we  were  able  to  differ- 
entiate the  curative  substance  of  1911,  but  only  when  we  undertook 
the  extraction  of  the  rice  polishings  ourselves.  When  this  was  done  in 
the  factory  no  curative  substance  was  obtained,  and  hence  no  pub- 
lication was  made,  thus  explaining  the  non-appearance  of  the 
protocols  of  the  animal  experiments.  Therefore,  it  is  not  justifiable 
to  apply  the  conclusions  drawn  from  our  negative  results  with  rice 
polishings  to  our  positive  yeast  findings,  which  we  shall  describe  in 
the  next  chapter. 

In  conclusion  we  wish  to  call  attention  to  an  investigation  by 
Hofmeister  and  Tanaka  (483).  The  impression  was  given  that  the 
active  vitamine  had  been  isolated  in  the  pure  state  from  rice  polish- 
ings. It  will  be  well  to  describe  Hofmeister 's  method,  different 
from  any  previously  used.  He  began  his  work  with  the  notion  that 
our  vitamine  is,  in  reality,  nicotinic  acid,  but  this  is  not  the  case, 
since  in  every  case  where  nicotinic  acid  was  isolated  there  was  no 
curative  action. 

Hofmeister  shook  rice  meal  three  times  with  double  the  volume  of 
80  per  cent  alcohol  on  the  shaking  machine.  The  filtered  solution 
was  evaporated  in  the  presence  of  a  stream  of  air,  in  vacuum.  Then 
the  residue  was  acidified  up  to  3  per  cent  with  hydrochloric  acid,  the 
fatty  acids  extracted  with  ether,  the  ether  removed,  the  solution 
concentrated  to  a  syrup  in  vacuum  at  a  low  temperature,  and  again 
taken  up  with  80  per  cent  alcohol  to  free  it  from  colloidal  impurities. 
The  clear  filtrate  was  made  faintly  alkaline  with  sodium  carbonate, 
taking  care  to  prevent  an  excess  of  alkali,  and  precipitated  by  bismuth 
potassium  iodide  (prepared  according  to  Kraut)  with  constant 
stirring.  It  is  necessary  here  to  avoid  a  strongly  acid  reaction,  else 
the  active  substance  precipitates  too.  The  dirty  grayish-yellow 
precipitate  of  the  choline  fraction  was  filtered  after  standing  for  5 
hours;  to  the  filtrate,  was  added  one-tenth  of  its  volume  of  20  per 
cent  hydrochloric  acid,  and  the  active  substance  precipitated  out 


176  THE   VITAMINES 

with  the  same  reagent.  The  powdery,  brick  to  scarlet  red  precipitate 
is  quickly  filtered  off  by  suction,  thoroughly  drained,  carefully  rubbed 
up  in  a  mortar  with  silver  carbonate,  and  immediately  filtered. 
The  weakly  alkaline  solution,  containing  silver,  is  acidified  at  once 
with  hydrochloric  acid,  filtered  free  from  silver  chloride  and  evapo- 
rated in  vacuum  almost  to  dryness.  The  residue  crystallizes  in  weakly 
colored,  radiating  deliquescent  crystals.  From  the  crude  hydro- 
chloride,  of  which  5  to  10  mgm.  was  sufficient  to  cure  a  pigeon 
in  24  hours,  and  which  retained  its  activity  for  8  to  10  days,  a  yellow 
precipitate  was  obtained  with  gold  chloride,  which  was  amorphous 
at  first  and  then  crystalline,  and  which  crystallized  from  water  in 
plates  and  flat  prisms.  The  quantity  of  aurate  obtained  corresponded 
evidently  to  the  amount  of  hydrochloride  used  (m.  p.  273.5°C.; 
which  on  further  re  crystallization  rose  to  277°  C.,  uncorrected). 
This  aurate  was  not  yet  entirely  pure  and  was,  therefore,  converted 
back  to  the  hydrochloride  and  recrystallized  from  water;  from  the 
purer  crystals,  a  preparation  was  obtained,  poor  in  ash,  which  melted 
at  240°C.  and  gave  the  formula  CsHnNC^.  HC1.  From  this  prepara- 
tion, the  pure  chlor-aurate  was  then  prepared  and  analyzed.  The 
free  base,  called  "Oridin,"  is  a  white,  powdery,  hygroscopic  substance, 
ea-ily  soluble  in  water  with  a  slight  acid  reaction;  it  is  slightly  soluble 
in  cold  absolute  alcohol  but  quite  so  in  hot.  From  the  latter,  the 
base  may  be  obtained  in  crystals  with  ether.  When  these  are  heated, 
pyridine-like  vapors  are  given  off.  In  water,  precipitates  were 
obtained  with  phosphotungstic  and  phosphomolybdic  acid,  bismuth 
potassium  iodide  and  gold  chloride,  but  not  with  bromine  water, 
platinic  chloride  or  sublimate.  The  phosphotungstic  acid  precipitate 
is  soluble  in  acetone,  but  not  very  soluble  in  water.  The  watery 
solution  does  not  dissolve  any  copper  carbonate  and  gives  no  color 
reaction  with  iron  salts.  Isonitrile  and  mustard  oil  reactions  were 
negative.  On  boiling  with  hydriodic  acid  and  phosphorus,  alkaline 
vapors  are  evolved,  but  not  with  sodium  hydroxide.  With  calcium 
hydrate  a  pyridine  odor  is  obtained.  The  purest  preparation  gave 
only  a  very  faint  pine  splinter  reaction.  No  color  was  obtained  with 
formaldehyde  and  sulphuric  acid,  or  on  heating  with  nitric  acid  and 
on  addition  of  alkali.  The  formula  is  isomeric  with  betaine,  and  the 
substance  is  perhaps  a  dioxypiperidine.  The  pure  substance  was 
inactive,  and  Hofmeister  seemed  uncertain  as  to  whether  the  active 
substance  had  been  destroyed  or  had  passed  into  the  filtrate.  We 


THE   ANTIBERIBERI   VITAMINE  177 

have  often  tried  to  precipitate  the  vitamine  with  gold  chloride  but 
without  succeeding.  Besides,  as  we  have  lately  again  noted,  a  num- 
ber of  substances  including  nicotinic  acid,  are  precipitated  by  bismuth 
potassium  iodide.  According  to  Hx>fmeister  (I.e.  4$3)  colamine, 
proline,  hematine,  papaverine,  trigonelline  and  stachydrine  have  no 
influence  on  beriberi  in  pigeons.  We  have  tried  the  above  pro- 
cedure with  tomatoes  but  found  it  impractical. 

CHEMICAL   INVESTIGATION   OF  YEAST 

The  chemical  aspect  of  vitamine  B  has  been  neglected  in  most  of 
the  books  and  reviews  on  the  subject.  We  propose  to  describe  only 
the  indisputable  facts  in  order  to  give  the  reader  the  opportunity  of 
forming  his  own  opinion  of  the  status  of  the  question.  It  is  too 
soon  to  say  whether  the  isolation  of  this  vitamine  has  been  successful 
or  not.  In  any  case,  the  investigation  on  yeast  extract  made  by  us 
is  one  of  the  few  researches  upon  which  we  may  build  further.  We 
have  investigated  yeast  in  various  ways,  and  yet  we  do  not  know  if 
the  vitamine  occurs  free  or  combined.  The  facts  uncovered  up  to 
the  present  point  to  both  possibilities. 

In  our  first  attempt  to  isolate  the  vitamine  from  yeast  (I.e.  324), 
75  kilos  of  dried  yeast  were  extracted  with  alcohol,  the  extract  con- 
centrated in  vacuum  and  the  residue  hydrolyzed  with  5  per  cent 
sulphuric  acid  for  five  hours;  after  cooling,  it  was  filtered  from 
separated  fatty  acids,  the  filtrate  diluted  with  an  equal  volume  of 
water  and  precipitated  with  phosphotungstic  acid.  The  precipitate 
was  decomposed  in  the  usual  manner,  and  the  resulting  filtrate  con- 
centrated. A  concentrated  watery  solution  of  silver  nitrate  was  then 
added  till  a  drop  of  the  clear  solution  gave,  with  a  saturated  baryta 
solution,  instead  of  a  white  precipitate,  a  brown  precipitate  of  silver 
oxide.  The  solution,  freed  from  the  heavy  precipitate  of  purine 
bases,  was  treated  with  saturated  baryta  so  long  as  it  gave  a  white 
precipitate  with  ammoniacal  silver  nitrate  solution.  The  silver 
precipitate  was  decomposed  with  hydrogen  sulphide  and  the  filtrate, 
after  the  removal  of  the  last  traces  of  baryta,  concentrated  to  small 
volume  in  vaccum;  the  residue  was  transferred  to  a  dish  with  a  little 
alcohol  and  allowed  to  stand  in  the  dessicator,  whereupon  crystals 
separated  out  weighing  0.6  gram.  They  were  re  crystallized  from 
dilute  alcohol  yielding  0.45  gram  of  colorless  needles  (see  fig.  40) 


178 


THE   VITAMINES 


melting  at  233°  C.  This  substance  from  yeast  was  therapeutically 
active;  2  to  4  centigrams  were  sufficient  to  cure  a  sick  pigeon  in  a 
short  time;  smaller  doses  were  not  tried.  The  substance  could  be 
precipitated  by  mercuric  acetate  but  not  by  the  nitrate  or  sulphate. 
It  gave  no  precipitate  with  a  nitron  solution — showing  the  absence 
of  nitric  acid — and  had  a  neutral  reaction.  No  blue  copper  salt 
was  obtained  with  copper  oxide,  which  would  indicate  that  the 
substance  is  not  an  amino-acid.  The  filtrate  from  the  silver  pre- 
cipitate was  inactive. 


FlG.  40.  MlCROPHOTOGRAPH  OF  CRYSTALS  OBTAINED  FROM  VlTAMINE  FRACTION 
OF  YEAST,  POSSESSING  CURATIVE   ACTION 

Another  batch  of  yeast  was  treated  in  a  somewhat  different  manner. 
The  alcoholic  extract,  after  concentration  in  vacuum,  was  simply 
taken  up  with  water,  and  not  hydrolyzed  as  above.  Thus  the 
vitamine  was  obtained  in  watery  solution.  This  was  then  pre- 
cipitated with  phosphotungstic  acid  and  treated  as  above;  in  this 
case  however,  the  silver  fraction  did  not  yield  an  active  product,  a 
substance  being  isolated  which  could  be  identified  as  uracil.  All  the 
vitamine,  in  this  case,  was  found  in  the  filtrate  from  the  silver  nitrate 
precipitate,  which  might  mean  that  the  vitamine  of  yeast  occurs 


THE    ANTIBERIBERI    VITAMINE 


179 


chiefly  in  combination.  This  filtrate  was  then  hydrolyzed  with 
sulphuric  acid,  and  the  subsequent  procedure  again  repeated.  Here 
too  we  met  with  a  mixture  of  pyrimidine  bases,  among  which  thymine 
could  be  demonstrated,  so  that  finally  after  a  long  procedure,  only  a 


FIG.  41.  PIGEON  BEFORE  TREATMENT 


FIG.  42.  THE  SAME  PIGEON  (CP.  FIG.  41)   CURED   WITH 
VITAMINE  AFTER  Two  HOURS 


MGM.    OF  YEAST 


trace  of  active  substance  was  found.  This  observation  shows  that 
in  working  with  vitamines,  the  number  of  manipulations  had  best 
be  kept  down  to  a  minimum,  if  results  are  to  be  obtained. 

Later  on  (485,  and  I.e.  465),  we  again  investigated  the  vitamine 
fraction,  using  2.5  grams  of  the  crude  fraction,  obtained  from  100 


180  THE   VITAMINES 

kilos  of  dried  yeast,  according  to  the  method  described.  The  sub- 
stance was  obtained  from  the  decomposition  of  the  silver  baryta 
fraction,  and  possessed  a  crystalline  structure,  melting  at  210°  C. 
(uncorrected) .  This  crude  vitamine  fraction  was  a  splendid  thera- 


FIG.  43.  PIGEON  BEFORE  TREATMENT 


FIG.   44.   SAME   PIGEON    (CF.    FIG.   43)  AFTER  THREE  HOURS,  WITH  4  MGM. 

OF  YEAST  VITAMINS 

peutic  agent  in  a  great  number  of  beriberi  pigeons.  As  the  accom- 
panying photograph  shows  (fig.  4 1-46),  the  preparation  cured  pigeons 
completely  in  a  very  short  time  (2  to  3  hours) .  Further  data  on  this 
subject  will  be  found  in  the  protocol  appearing  at  the  end  of  this  chap- 
ter, showing,  in  the  form  of  a  table  (p.  185),  the  action  of  substances 


THE    ANTIBERIBERI   VITAMINE 


181 


obtained  from  yeast.  The  dose  of  crude  fraction  used  was  4  to  8 
mgm.,  and  the  preparation  was  administered  intramuscularly.  The 
product  gave  a  strong  uric  acid  and  phenol  reaction  with  the  Folin- 
Macallum  reagents  (486). 


FIG.  45.  ^PIGEON  BEFORE  TREATMENT 


FIG.  46.  SAME  PIGEON  (CF.  FIG.  45)   AFTER   THREE   HOURS,   WITH  8  MGM. 

YEAST  VITAMINE 

This  2.5  grams  of  crude  fraction  was  then  recrystallized  from 
dilute  alcohol,  and  gave,  at  first,  1.6  grams  of  a  substance  crystallizing 
in  microscopic  needles.  Even  after  several  recrystallizations,  the 
melting  point  could  not  be  raised  above  229°  C.  (corrected).  The 
first  crystallization  still  gave  the  reactions  mentioned  above;  only 


182  THE    VITAMINES 

when  the  substance  was  recrystallized  twice  did  these  reactions  dis- 
appear. The  purified  substance  was  compared  with  that  obtained 
before,  and  mixed  melting  points  were  taken,  whereby  the  new 
product  melted  four  degrees  lower.  Mixed  in  equal  quantities,  both 
substances  melted  at  229°C.  (corrected).  They  were  absolutely 
identical  as  regards  solubility,  reactions  and  crystalline  form.  The 
substance  gave  a  white  precipitate  with  Millon's  reagent,  but  the 
color  reactions  were  always  negative.  In  a  0.74  per  cent  solution, 
a  definite  rotation  in  the  polarimeter  could  not  be  shown.  On 
drying  in  vacuum  at  100°  C.  and  analyzing,  the  following  figures 
were  obtained: 

3.267  mgm.  yielded  6.63  mgm.  CO2  and  1.05  mgm.  H2O;  55.35  per  cent  C,  3.60 

per  cent  H. 
4.224  mgm.  yielded  8.425  mgm.  CO2  and  1.36  mgm.  H2O;  55.72  per  cent  C,  3.69 

per  cent  H. 
4.256  mgm.  yielded  8.66  mgm.  CO2  and  1.36  mgm.  H2O;  55.50  per  cent  C,  3.58 

per  cent  H. 

3.048  mgm.  yielded  0.373  cc.  N.  (710  mm.  15.5°C.)  13.53  per  cent  N. 
3.627  mgm.  yielded  0.430  cc.  N  (712  mm.;  14°C.)  13.21  per  cent  N. 
3.286  mgm.  yielded  0.401  cc.  N  (705  mm.;  18°C.)  13.28  per  cent  N. 

These  figures  correspond  to  the  formula  C^HigOg^  (521.24), 
which  gives  55.25  per  cent  C,  3.68  per  cent  H  and  13.44  per  cent  N. 
Since  a  definite  crystalline  substance  is  noted  on  the  tube  when  drying 
at  100°  C.  in  vacuum,  analyses  were  made  after  the  substance  was 
dried  in  vacuum  over  sulphuric  acid;  the  following  figures  were 
obtained : 

3.733  mgm.  gave  7.775  mgm.  CO2  and  1.185  mgm.  H2O;  56.80  per  cent  C,  3.55 

per  cent  H. 
3.538  mgm.  gave  7.35  mgm.  CO2  and  1.195  mgm.  H2O;  56.66  per  cent  C,  3.78 

per  cent  H. 
3.174  mgm.  gave  0.372  cc.  N  (702  mm.;  17°C.)  12.74  per  cent  N. 

These  figures  correspond  to  the  formula  C26H2i09N5,  which  gives 
57.01  per  cent  C,  31.87  per  cent  H  and  12.81  per  cent  N.  From  this, 
it  appears  that  on  drying  at  higher  temperatures  a  substance  rich  in 
carbon  is  lost.  This  characteristic  and  the  great  difference  in 
elementary  composition  excludes  the  possibility  that  this  substance 
was  nicotinic  acid.  We  have  recently  attempted  to  secure  further 
evidence  in  this  connection  by  the  use  of  the  yeast  method.  If  the 
nitrogen  in  this  substance  is  estimated  by  the  method  of 


THE   ANTIBERIBEKI   VITAMINE  183 

Kjeldahl,  only  a  third  of  it  is  determined.  0.1328  gram  needed 
4.0  cc.  ye-  H2S04;  found,  4.21  per  cent  N.  If  the  substance  is 
titrated  with  sodium  hydroxide  against  phenolphthalein,  then 
0.0373  gram  requires  2.8  cc.  •&  NaOH.  For  a  four  basic  acid 
of  the  formula  C24Hi9O9N5,  the  calculated  amount  is  2.8  cc.  That 
this  substance  is  of  an  acid  nature,  is  very  important  and  we  shall 
consider  this  phase  of  the  matter  again.  As  regards  the  therapeutic 
action  of  substance  I,  examples  are  given  in  the  table  (p.  185). 

The  filtrate  and  mother  liquor  combined,  of  substance  I,  which 
gave  a  strong  phenol  and  uric  acid  reaction,  were  precipitated  by 
picric  acid;  a  crystalline  precipitate  was  obtained,  amounting  to  0.9 
gram;  which  was  first  recrystallized  from  a  mixture  of  acetone  and 
alcohol  and  then  from  water.  The  once  recrystallized  substance  still 
gave  a  positive  phosphotungstic  acid  reaction.  On  purification  (light 
yellow  needles),  the  melting  point  was  raised  from  217°  to  219°  C. 
(uncorrected)  and  gave  the  following  figures  on  combustion: 

4.70  mgm.  gave  7.115  mgm.  CO2  and  0.98  mg.  H2O;  41.29  per  cent  C;  2.33  per 

cent  H. 

2.966  mgm.  gave  0.436  cc.  N  (708  mm.,  19.5°C.),  15.99  per  cent  N. 
4.212  mg.  gave  0.61  cc.  N  (711  mm.,  18°C.),  15.89  per  cent  N. 
0.1061  grain  gave  0.1652  Nitron  picrate,  65.91  per  cent  picric  acid. 
0.1261  grain  gave  0.1951  Nitron  picrate,  65.49  per  cent  picric  acid. 

Calculated  for  Ci2H809N4  (352. 12) -40.9  per  cent  C,  2.29  per  cent 
H,  15.91  per  cent  N  and  65.05  per  cent  picric  acid.  This  picrate  was 
quantitatively  decomposed  with  nitron,  and  on  concentrating  gave 
0.45  gram  of  silky,  lustrous,  colorless  needles,  the  melting  point  of 
which,  after  recrystallization,  rose  to  235°  C.  (uncorrected).  The 
crystals  were  very  easily  soluble  in  water  and  gave  no  color  reactions. 
When  dried  at  100°C,  in  vacuum,  the  analysis  gave  the  following 
figures : 

3.432  mgm.  yielded  7.345  mgm.  CO2and  1.10  mgm.  H2O;  58.37  per  cent  C;3.62 

per  cent  H. 
3.784  mgm.  yielded  8.11  mgm.  CO2  and  1,325  mgm.  H2O;  58.45  per  cent  C. 

3.92  per  cent  H. 

3.415  mgm.  yielded  0.343  cc.  N.  (702  mm.,  18.5°C),  10.87  per  cent  N. 
2.562  mgm.  yielded  0.252  cc.  N  (15.5°C.),  10.92  per  cent  N. 

Calculated  for  C6H502N  (123.05) -58.3  per  cent  C,  41.08  per  cent 
H  and  11.34  per  cent  N.  The  substance  was  evidently  nicotinic  acid. 


184  THE   VITAMINES 

Mixed  with  the  nicotinic  acid  from  rice  polishings,  it  showed  the  same 
melting  point,  and  the  picrates  behaved  similarly. 

The  mother  liquor  of  the  picrate  precipitate,  which  gave  a  strong 
uric  acid  and  phenol  reaction,  was  shaken  out  in  acid  solution  with 
ether  in  order  to  remove  the  picric  acid.  After  elimination  of  the 
hydrochloric  acid,  the  solution  was  slowly  evaporated,  yielding 
0.4  gram  of  a  substance  melting  at  210°  C.  and  when  recrystallized 
from  dilute  alcohol  to  constant  melting  point,  melted  at  222  to  223°  C. 
(microscopic  needles,  somewhat  more  soluble  in  water  than  substance 
I) .  The  pure  substance  no  longer  gave  the  uric  acid  nor  the  Millon 
reaction,  but  the  phenol  reaction  was  still  positive.  For  analysis, 
one  sample  was  dried  in  the  vacuum  dessicator  over  sulphuric  acid  at 
ordinary  temperature;  another  was  dried  in  vacuum  at  100°  C.  and 
another,  at  115°  C.  The  substance  dried  over  sulphuric  acid  gave 
the  following  figures: 

3.559  mgm.  gave  7.745  mgm.  CO2  and  1.21  mgm.  H20;  59.35  per  cent  C;  3.80 

per  cent  H. 
3.445  mgm.  gave  0.375  cc.  N  (706  mm.  17°C.),  11.90  per  cent  N. 

Dried  in  vacuum  at  100°C. 
4.172  mgm.  gave  9.065  mgm.  CO2  and  1.535  mgm.  H2O;  59.26  per  cent  C;  4.12 

per  cent  H. 
2.716  mgm.  gave  0.295  cc.  N  and  (711  mm.  15°C.)  12.04  per  cent  N. 

Dried  in  vacuum  at  115°C. 
3.694  mgm.  gave  7.99  mgm.  CO2  and  1.22  mgm.  H20;  58.99  per  cent  C;  3.70  per 

cent  H. 

These  figures  correspond  equally  well  to  two  formulas,  though 
somewhat  better  to  the  second. 


CALCULATED  FOR 

C2sHi807N4  (462.22) 

C29H23O9N5(585.28) 

C  . 

per  cent 

59.71 
3.84 
12.12 

per  cent 

59.46 
3.96 
11.97 

H 

N  

To  recapitulate  briefly  the  yeast  fractionation,  we  have  been  able 
to  isolate  three  different  substances  from  the  vitamine  fraction, 
which  gives  a  strong  uric  acid  and  phenol  reaction. 


THE    ANTIBERIBERI    VITAMINE 


185 


SUBSTANCE 

MELTING 
POINT 

URIC  ACID 
REACTION 

PHENOL 
REACTION 

REMARK 

C24H:9O9N5  (dried  at  100°C) 
C26H2iO9N5   (dried  at  ordi- 
nary temperature)  
C^HaaOaNa 

°c. 
229 

223 
222-223 

- 

+ 

Gives  no  picrate 

Gives  no  picrate 
Gives  no  picrate 

C6H5O2N  

235 

Gives       picrate 

Nicotinic  acid 

Aside  from  this,  the  first  substance,  on  heating  in  vacuum,  loses 
SL  small  amount  of  substance  rich  in  carbon,  giving  lower  analytical 
results.  Although  the  statement  is  often  made  that  we  had  obtained 
only  impure  nicotinic  acid,  it  is  not  justified,  since  the  analytical 
figures  are  too  far  apart  from  the  composition  and  the  characteristics 
of  this  acid.  Regarding  the  therapeutic  action  of  these  substances, 
many  experiments  were  performed,  some  of  which  naturally  turned 
out  negative.  In  the  table,  the  product  C26H2i09N5  is  called 
substance  I,  and  C^H^OgNs,  substance  II. 


NUM- 
BER OF 
ANI- 
MALS 

SUBSTANCE 

DOSE 

ACTION 

DATS 
SUR- 
VIVED 

mgm. 

1 

Crude  crystallization 

4 

Cured  in  3  hours 

4 

2 

Crude  (fig.  41-42) 

8 

Cured  in  2  hours 

6 

3 

Crude  (fig.  43-44) 

4 

Cured  in  3  hours 

4 

4 

Crude  (fig.  45-46) 

8 

Cured  in  3  hours 

4 

5 

Substance  I 

8 

Partially   cured  in  7 

hours. 

3 

6 

Substance  I 

4 

Improvement 

4 

7 

Substance  I 

2 

Cured  in  a  few  hours 

3 

8 

Substance  I  +  Nicotinic  acid 

5+2 

Cured  in  2^  hours 

4 

9 

Substance  I  +  Nicotinic  acid 

4+2 

Cured  in  3     hours 

5 

10 

Substance  I  +  Nicotinic  acid 

3+2 

Cured  in  2     hours 

6 

11 

Substance  I  +  Nicotinic  acid 

4+2 

Cured  in  2^  hours 

7 

12 

Substance  I  +  Nicotinic  acid 

3+2 

Cured  in  4     hours 

4 

13 

Nicotinic  acid 

10 

Negative 

2 

14 

Nicotinic  acid 

5 

Slight 

3 

15 

Nicotinic  acid 

4 

Negative 

2 

16 

Nicotinic  acid 

4 

Negative 

1 

17 

Substance  II 

5 

Negative 

— 

IS 

Substance  II 

5 

Negative 

1 

186  THE   VITAMINES 

All  these  substances  were  administered  to  the  pigeons  intra- 
muscularly, and  from  the  results,  it  is  evident  that  substance  I  had 
the  greatest  therapeutic  action. 

Others  also  investigated  this  question.  Barsickow  (487)  concluded 
that  the  therapeutic  action  was  inherent  only  in  the  living  or  dead 
yeast  cells,  and  for  the  most  part,  he  found  the  extracts  inactive; 
this  has,  of  course,  been  shown  to  be  incorrect.  Edie,  Evans,  Moore, 
Simpson  and  Webster  (488)  did  not  hydrolyze  the  yeast,  but  treated 
the  alcoholic  extract,  after  the  concentration  of  the  solvent,  directly 
with  silver  nitrate  and  baryta.  They  obtained  in  this  way  a  hygro- 
scopic syrup  which  was  very  active  for  pigeons,  6  mg.  being  suffi- 
cient, as  a  rule,  for  a  cure.  On  further  purification,  a  crystalline 
substance  was  obtained  which,  because  of  the  small  yield,  could  not 
be  purified  further.  This  crystalline  substance  was  analyzed  but  the 
data  on  its  therapeutic  action  were  lacking. 

Another  method,  useful  for  the  purpose  of  removing  a  large  mass  of 
inactive  material,  was  described  by  us  (489)  in  1916.  This  method  is 
based  upon  the  separation  of  the  phosphotungstates  by  means  of 
the  varying  solubility  in  acetone.  The  residue,  obtained  from  the 
alcoholic  extract  of  100  kilos  of  dried  yeast,  was  extracted  with  10 
per  cent  sulphuric  acid,  this  extract  was  then  diluted  half  again  with 
water  and  precipitated  with  phosphotungstic  acid.  The  resulting 
precipitate,  weighing  2237  grams  was  triturated  several  times  with 
dry  acetone,  leaving  in  the  end  144  grams  of  insoluble  material.  This 
portion,  which  represents  5.1  per  cent  of  the  total  precipitate,  con- 
tained all  the  vitamine,  according  to  our  animal  experiments.  The 
precipitate  was  decomposed  with  neutral  lead  acetate2  and  the 
resulting  solution  precipitated  with  picric  acid.  The  voluminous  pre- 
cipitate consisting  of  adenine  picrate  was  filtered  off.  It  was  impos- 
sible to  crystallize  the  vitamine  from  the  mother  liquor,  making  it 
necessary  to  continue  the  fractionation  with  sublimate,  platinic 
chloride,  and  picrolonic  acid.  The  course  of  the  fractionation,  which 
was  controlled  by  animal  experiments,  is  shown  in  the  diagram 
below,  in  which  the  double  lines  indicate  the  fraction  containing  the 
vitamine.  According  to  a  patent  by  Bohringer  and  Sons  (491)  the 

2  We  also  attempted  to  decompose  the  precipitate  by  shaking  it  out  with 
amyl  alcohol  in  acid  solution  according  to  Van  Slyke  (490),  but  the  method 
was  not  quantitative  because  the  last  traces  of  phosphotungstic  acid  are 
removable  only  with  great  difficulty. 


THE    ANTIBERIBERI    VITAMINE  187 

Phosphotungstic  acid  precipitate  from  yeast. 


II  I 

Acetone  insoluble  part  Acetone  soluble  part 


/  II 

Picrate  Picric  acid  nitrate 

II 

ir  \ 

Sublimate  precipitate  Sublimate  filtrate 


Platinic  chloride  precipitate      l| 

Filtrate 


Sublimate  precipitate  Sublimate  filtrate 

phosphotungstic  acid  in  the  above  method  was  substituted  by  phos- 
photitanic  acid.  Cooper  (492)  then  showed  that  when  pressed  yeast 
is  allowed  to  stand  for  35  hours  at  35°  C.,  the  mass  becomes 
semi-liquid  and  on  filtration  from  the  cell  residue,  a  liquid  called 
"autolyzed  yeast"  is  obtained,  which  contains  almost  all  of  the 
vitamine  of  the  yeast.  It  was  shown  later  that  the  yeast  con- 
stituents are  not  completely  autolyzed,  since  if  the  liquid  is  heated 
to  60  to  70°  C.,  an  appreciable  amount  of  protein  separates  out. 
When  the  acetone  method  was  applied  to  this  liquid,  the  amount  of 
the  acetone  insoluble  portion  was  much  larger — about  1125  grams 
from  5  kilos  of  pressed  yeast.  We  then  showed  that  when  the 
phosphotungstates  of  choline,  betaine,  stachydrine,  guanine,  adenine, 
guanidine,  creatinine  and  nicotinic  acid  were  tested  for  their 
solubility  in  acetone-water  solutions  of  various  dilutions,  the  sol- 
ubility was  as  follows : 

25  per  cent  acetone     — >     100  per  cent     — >     50  per  cent-    — >     75  per  cent 

Choosing  these  four  concentrations,  the  solubility  was  least  in  25 
per  cent  acetone  and  greatest  in  75  per  cent  acetone.  We  were  then 
able  to  show,  in  our  work  with  Dubin  (493),  that  autolyzed  yeast 
contains  much  more  vitamine  than  vitamine  extracts  obtained  from 
yeast  by  various  solvents.  For  this  reason,  autolyzed  yeast  would 
be  superior  to  all  other  extracts  for  the  purpose  of  vitamine  fraction- 


188  THE   VITAMINES 

ation,  if  a  large  quantity  of  inactive  extractives  did  not  go  into 
solution  at  the  same  time.  If  the  phosphotungstic  acid  precipitate 
from  autolyzed  yeast  is  treated  directly  with  dry  acetone,  almost  a 
third  of  it  remains  insoluble,  including  a  large  amount  of  purine 
derivatives.  If  the  acetone-water  method  is  used,  as  outlined  above, 
we  obtain  from  the  above-mentioned  precipitate,  weighing  1125 
grams,  the  following  fractions : 


ACETONE  CONCENTRATION 

AMOUNT  SOLUBLE 

per  cent 

grams 

per  cent 

25 

522 

46.4 

100 

190 

16.8 

50 

155 

13.7 

.      75 

108 

9.6 

Total  precipitate — 1125  grams. 

Insoluble  residue— 150  grams  =  13.3  per  cent. 

This  method  might  be  of  importance  but  in  this  case,  no  animal 
experiments  were  carried  out.3 

A  method  indicative  of  great  progress  at  the  time  was  described 
by  Seidell  (495).  If,  for  example,  1  liter  of  autolyzed  yeast  is  shaken 
with  50  grams  of  Lloyd's  reagent  (fuller's  earth),  allowed  to  stand  for 
several  hours  and  the  residue  filtered  off  and  washed  with  dilute 
hydrochloric  acid,  vitamine  B  is  quantitatively  removed  from  the 
solution,  and  the  filtrate  is  completely  inactive.  The  great  hopes 
held  out  for  this  method,  however,  have  not  materialized.  In  the 
first  place,  it  is  very  difficult  to  remove  the  vitamine  from  the  fuller's 
earth,  and  secondly,  a  number  of  other  substances,  difficult  to  remove, 
are  adsorbed  together  with  the  vitamine.  In  the  end,  R.  R.  Williams 
and  Seidell  (496)  were  able  to  extract  the  vitamine  from  the  fuller's 
earth  with  5  per  cent  sodium  hydroxide,  at  the  same  time  showing 
that  the  vitamine  was  contaminated  with  considerable  adenine. 
Osborne  and  Leavenworth  (496a)  showed  that  dilute  alkali  has 
no  destructive  action  of  vitamines,  especially  if  the  contact  is  of 
short  duration.  For  liberation  of  vitamine  from  activated  fuller's 
earth  all  those  methods  are  suitable  which  are  used  in  freeing  alkaloids 

3  In  the  meantime,  this  method  was  elaborated  by  Drummond  (494)  in 
that  he  investigated  the  solubility  of  various  phosphotungstates,  and  dis- 
cussed the  practical  applicability  of  these  data. 


THE    ANTIBERIBERI    VITAMINE  189 

from  fuller's  earth  preparations  by  Rhodehamel  and  Stuart  (496b). 
Recently,  Seidell  (496c)  has  combined  the  fuller's  earth  method 
with  our  silver  method  and  has  obtained  some  promising  results. 
He  liberates  the  vitamine  with  a  baryta  solution,  the  procedure 
being  hastened  as  much  as  possible.  From  the  nitrate,  the  purine 
bases  are  eliminated  with  silver  nitrate  while  the  vitamines  are 
precipitated  by  ammoniacal  silver  nitrate.  This  fraction  can  be 
reprecipitated  in  the  same  way  and  can  be  analyzed  for  its  silver 
content.  A  crystalline  substance  was  isolated  from  this  fraction, 
but  it  proved  inactive,  showing  it  to  be  a  fairly  complicated  mixture. 
We  found  that  nicotinic  acid  is  also  precipitated  by  ammoniacal 
silver  nitrate.  For  the  liberation  of  vitamine  from  activated  fuller's 
earth,  we  have  likewise  used  ammonia  and  pyridine;  we  were  also 
able  to  confirm  the  findings  of  Eddy,  Heft,  Stevenson  and  Johnson 
(I.e.  129e)  that  vitamine  can  be  adsorbed  with  a  specially  pre- 
pared charcoal  from  which  it  can  be  liberated  with  glacial  acetic 
acid.  Voegtlin  and  White  (497)  adsorbed  the  vitamine  with  mastic, 
and  with  colloidal  arsenic  sulfide,  in  which  case  the  impurity  was 
guanine  instead  of  adenine. 

Drummond  (498)  attempted  to  confirm  our  findings  on  vitamine 
B,  and  although  he  was  able  to  do  so  for  the  most  part,  he  concluded 
that  our  chemical  work  was  of  no  particular  significance,  because  he 
held  that  the  vitamines  were  not  precipitated  by  the  various  pre- 
cipitation methods,  but  were  adsorbed  by  the  precipitates.  The 
characteristic,  adsorbability,  is  peculiar  to  many  pure  chemical  sub- 
stances. We  need  only  recall  Lloyd's  reagent  which,  as  we  have 
personally  proved  by  tests  on  rats,  quantitatively  adsorbs  strychnine 
and  other  alkaloids.  Pure  glucose  is  also  adsorbed  in  very  appre- 
ciable amounts  by  various  kinds  of  blood  charcoals.  Besides,  Drum- 
mond carried  out  the  silver  nitrate-baryta  precipitation  in  a  manner 
different  from  ours,  and  used  rats  as  experimental  animals,  which, 
as  we  shall  see,  behave  differently  than  do  pigeons  and  chickens. 

Abderhalden  and  Schaumann  (499)  have  recently  published  con- 
siderable work  on  yeast,  which  unfortunately  is  described  so 
unclearly  that  it  is  hard  to  state  whether  or  not  progress  has  been 
made.  These  authors  declare,  in  accord  with  the  phosphorus  theory 
of  Schaumann,  that  vitamine  B  of  yeast  occurs  as  a  nucleoproteid. 
They  were  able  to  isolate  an  active  nucleoproteid  from  yeast. 
Pigeons  receiving  a  half  gram  of  this  substance  daily  lived  for  70 


190  THE    VITAMINES 

days.  We  have  already  seen  that  such  findings  possess  no  special 
significance,  since  the  activity  of  the  nucleoproteid  might  have 
been  due  to  adsorption,  as  with  fullers  earth.  The  authors  also 
assume  that  the  mother  substance  of  the  vitamine  varies  according 
to  the  source  of  material.  The  vitamine  was  obtained  from  yeast 
extracts  by  precipitation  with  acetone,  and  purified  by  several 
reprecipitations.  This  crude  fraction  was  investigated  physiologi- 
cally, and  found  curative  for  beriberi,  but  it  could  not  keep  pigeons 
alive  for  any  prolonged  period.  Another  preparation  was  obtained 
from  yeast  by  treatment  with  alkalis.  This  substance  did  not  cure 
beriberi,  but  when  added  together  with  the  acetone  preparation,  its 
effect  was  to  prolong  life.  A  number  of  substances  were  isolated  from 
the  acetone  preparation,  by  reprecipitating  with  sublimate,  among 
which  there  was  apparently  only  a  single  new  substance  obtained  in 
the  pure  state.  This  base,  called  "Aschamine"  had  the  structure  of 
dimethylpropenylamine,  the  formula  being 

CH  =  CH  •  CH2 

/ 
C5H130  N  =  OH— N— CH3 

\ 

CH3 
H 

it  was  inactive  for  beriberi.  In  the  other  fractions,  betaine  was 
found  as  an  impurity.  All  of  these  substances  were  prepared  from 
hydrolyzed  yeast,  and  the  authors  believed  that  the  active  substance 
belonged  to  the  betaine  group,  having  its  characteristic  structure. 
In  a  later  experiment,  Abderhalden  (500)  states  that  the  active 
substance  cannot  be  quantitatively  removed  from  yeast  by  absolute 
alcohol  or  absolute  alcohol  and  acetone. 

Sugiura  (501)  placed  yeast  and  also  carrot  extracts  in  collodion 
sacs,  and  permitted  the  contents  to  dialyze  through  the  membrane, 
whereupon  the  product  crystallized  on  the  outside  of  the  membrane. 
It  was  evidently  hoped  to  prepare  the  active  substance  in  the  pure 
state  by  this  simple  procedure,  but  naturally  the  entire  yeast  mixture 
dialyzed  out. 

Osborne  and  Wakeman  (502)  described  a  method  which  promised 
greater  success.  The  procedure  consisted  chiefly  in  eliminating 
the  autolysis  of  the  yeast  cells  by  washing  them  and  boiling  with 
water,  to  which  some  acetic  acid  has  been  added,  thus  coagulating 


THE    ANTIBERIBERI    VITAMINE  191 

the  protein  and  obtaining  an  extract  free  from  the  products  of  auto- 
lysis.  Concretely,  4.5  kilos  of  fresh  yeast,  corresponding  to  830 
grams  of  dry  yeast,  was  crumbled  into  boiling  water,  the  mass  centri- 
fuged,  the  residue  washed  with  water  and  again  centrifuged.  Alto- 
gether, 15  liters  of  water  were  used,  the  combined  extracts  were 
evaporated  to  two  liters  and  poured  into  three  liters  of  93  per  cent 
alcohol,  giving  a  53  per  cent  alcoholic  content  by  weight.  The  pre- 
cipitate, which  was  inactive,  was  filtered  off  and  the  filtrate,  together 
with  the  washings,  evaporated  to  300  cc.  This  liquid  was  poured 
into  1960  cc.  of  93  per  cent  alcohol,  giving  a  79  per  cent  alcoholic 
content.  The  second  precipitate  was  dissolved  twice  in  water  and 
precipitated  with  alcohol  so  that  the  alcoholic  content  should  be 
90  per  cent.  This  fraction  was  supposed  to  contain  almost  the 
entire  quantity  of  vitamine  B  present  in  the  yeast,  and  amounted  to 
6.2  per  cent  of  the  dry  yeast.  The  preparation  had  an  acid  reaction 
and  gave  a  heavy  precipitate  with  lead  acetate  and  sublimate;  25 
per  cent  of  this  fraction  could  be  precipitated  with  silver  nitrate - 
baryta.  This  work  shows  that  the  vitamine  of  non-hydrolyzed  yeast 
is  insoluble  in  strong  alcohol.  In  our  experiments  this  method  did 
riot  prove  successful,  as  in  each  fresh  sample  of  yeast  the  limits  of 
precipitation  have  to  be  established  anew. 

There  is  also  the  report  on  the  chemistry  of  yeast  by  Myers  and 
Voegtlin  (503).  They  showed  that  vitamine  B  could  be  extracted 
from  yeast  by  shaking  the  latter  with  olive  oil;  the  procedure  obvi- 
ously possesses  no  particular  advantage  over  the  usual  extraction,  so 
that  later  they  used  methyl  alcohol  acidified  with  hydrochloric  acid 
for  extraction.  The  resulting  extract  was  concentrated  in  vacuum 
and  the  residue  extracted  several  times  with  dilute  hydrochloric  acid. 
In  this  extract,  the  purine  bases  were  precipitated  with  silver  acetate. 
The  precipitate  was  filtered  off  and  more  silver  acetate  added  to  the 
filtrate,  followed  by  a  saturated  solution  of  baryta.  This  precipitate 
was  decomposed  in  the  usual  manner,  and  the  histidine  removed  from 
the  filtrate  with  mercuric  sulphate.  The  vitamine  was  then  pre- 
cipitated from  the  filtrate  with  absolute  alcohol.  The  precipitate 
was  dissolved  in  water  and  freed  from  mercury,  whereupon  a  very 
active  solution  was  obtained.  This  solution  gave  a  brown  diazo 
reaction  and  contained  a  product  similar  to  histamine.  When  the 
solution  was  concentrated,  crystals  were  obtained  which  were  active 
only  so  long  as  they  contained  some  mother  liquor.  On  washing  the 


192  THE    VITAMINES 

crystals  with  alcohol,  the  crystalline  form  changed  from  spindles  to 
prisms.  If  these  prisms  were  recrystallized  from  water,  spindles 
were  again  obtained.  In  this  experiment,  the  vitamine  was  inac- 
tivated by  drying.  The  adsorption  methods  were  shown  to  be 
unspecific. 

OTHER   SOURCES   OF  SUPPLY 

Hulshoff  Pol  (I.e.  59  and  504)  reported  further  experiments  with 
theX-acid  of  Katjang  idjoe beans  (Phaseolus  radiatus)  without  giving 
more  details  on  the  chemistry  of  the  substance.  In  1912,  we  inves- 
tigated milk  (I.e.  324)  with  this  point  in  view.  As  a  source  of  supply, 
we  used  a  dried  milk  preparation,  sold  under  the  name  "Trumilk." 
This  was  extracted  with  alcohol  and  ether.  The  combined  extracts 
were  concentrated  and  the  residue  hydrolyzed  with  10  per  cent 
sulphuric  acid  for  5  hours.  In  this  way,  51  grams  of  phosphotungstic 
acid  precipitate  were  obtained  from  1398  grams  of  dried  milk.  The 
silver-baryta  fraction  was  prepared  from  this  precipitate  in  the  usual 
manner.  The  decomposed  precipitate  yielded  a  small  amount  of 
crystals  melting  at  230°  C.,  and  was  curative  for  pigeon  beriberi.  In 
the  same  way,  it  was  possible  to  prepare  from  2180  grams  dried  ox 
brain  (I.e.  324)  220  grams  of  phosphotungstic  acid  precipitate,  and 
to  obtain  a  trace  of  a  crystalline  substance  from  the  silver-baryta 
fraction,  melting  at  203°  C.  and  having  curative  properties.  Voegtlin 
and  Towles  (505)  investigated  extracts  of  spinal  cord  and  found  that 
autolyzed  extracts  were  more  active  than  natural  extracts. 

From  42  liters  of  commercial  lime  juice  (I.e.  324),  we  obtained  1200 
grams  phosphotungstates,  from  which  there  resulted  5.9  grams  of  a 
vitamine  fraction.  Although  this  fraction  did  not  crystallize  out,  the 
solution  was  very  curative  for  pigeon  beriberi.  From  our  informa- 
tion of  the  vitamines  in  the  above  mentioned  publication,  the 
presence  of  vitamine  B  could  indeed  be  demonstrated,  and  yet  the 
crystalline  substance  itself  may  perhaps  have  been  impure  nicotonic 
acid  contaminated  with  vitamine. 

Sullivan  and  Voegtlin  (506)  fractionated  wheat  chaff  and  extracts 
of  peas  and  ox  liver.  The  residues  from  the  respective  alcoholic 
extracts  were  hydrolyzed  with  5  to  10  per  cent  sulphuric  acid  for  five 
hours  in  a  stream  of  CO2.  The  precipitates  obtained  with  phospho- 
tungstic acid  were  then  decomposed,  either  directly  or  after  dissolving 
in  50  per  cent  alcohol,  with  neutral  lead  acetate.  After  the  elimina- 


THE    ANTIBERIBERI    VITAMINE  193 

tion  of  lead,  the  filtrate  was  precipitated  by  an  excess  of  silver  acetate 
and  treated  with  baryta,  after  removal  of  purine  precipitates. 

Steenbock  (507)  prepared  from  egg-yolk  a  substance  soluble  in 
water-acetone,  active  and  not  quantitatively  precipitated  by  phospho- 
tungstic  acid.  McCollum  and  Simmonds  (I.e.  315)  sought  to  con- 
centrate vitamine  B  by  extraction  with  various  organic  solvents. 
They  found  that  beans  could  be  extracted  with  ether,  benzol,  or 
acetone,  without  dissolving  the  vitamine.  Subsequently  it  can  be 
extracted  with  alcohol  and  the  solution,  when  concentrated,  may  be 
taken  up  by  benzol.  The  same  method  was  applied  to  wheat  germ 
and  pigs'  liver. 

SYNTHETIC   EXPERIMENTS 

Although  we  undertook  some  synthetic  experiments  with  nicotinic 
acid,  based  on  its  occurrence  in  the  vitamine  fractions  obtained  from 
various  sources,  we  perceived  that  theoretical  reasons  for  this  trend  of 
thought  were  hardly  justifiable.  Nevertheless,  Williams  (I.e.  473), 
instituted  such  experiments.  He  condensed  p-oxy  nicotinic  acid  with 
itself  or  with  nicotinic  acid,  and  believed  that  the  first  condensation 
product  was  much  more  active  for  avian  beriberi  than  all  other 
similarly  synthesized  condensation  products.  Thereupon,  Williams 
(508)  proposed  the  very  attractive  theory  that  oxy-pyridine  and 
pyridine  carbonic  acid  occur  in  two  isomers,  which  possess  different 
crystalline  forms,  and  which  may  be  transformed  into  each  other 
under  certain  conditions.  The  labile  form  remained  intact  for  some 
days  and  in  this  case  was  therapeutically  active ;  then  it  changed  into 
the  inactive  stable  form.  Later,  Williams  (509)  extended  his  theory 
to  include  such  pyrimidine  and  purine  derivatives  as  were  theoreti- 
cally capable  of  forming  the  betaine  ring.  Williams  and  Seidell  (I.e. 
496)  tried  to  convert  adenine  obtained  from  autolyzed  yeast  by  ad- 
sorption with  fuller's  earth,  by  boiling  with  acetic  acid  and  acetic  anhy- 
dride, into  an  isomeric  form.  This  labile  form  was  supposed  to  have 
a  therapeutic  action;  after  re  crystallization,  this  peculiarity  was  sup- 
posed to  disappear,  the  substance  being  changed  back  again  to  the 
usual  adenine.  The  theory  of  Williams  explained  the  slight  stability 
of  vitamine  B  very  beautifully,  but  unfortunately  it  could  not  be  con- 
firmed. Voegtlin  and  White  (I.e.  497)  sought  in  vain  to  produce  an 
active  product  from  adenine,  while  Harden  and  Zilva  (510)  tried  to 
do  the  same  with  o-oxypyridine  and  adenine.  Since  the  course  of 


194  THE   VITAMINES 

reasoning  pursued  by  Williams  could  not  be  confirmed,  and  since 
he  has  published  no  further  related  work  since  1916,  the  subject  may 
apparently  be  considered  as  closed. 

STABILITY   OF   THE   ANTIBERIBERI   VITAMINE   AGAINST  HEAT  AND 
CHEMICAL   AND   PHYSICAL  AGENTS 

As  far  back  as  the  work  of  Grijns  (I.e.  52)  we  knew  that  the 
vitamine  B  from  Katjang-idjoe  beans  lost  its  curative  properties  on 
heating  to  120°  C.  Ever  since,  a  number  of  investigators  have 
occupied  themselves  with  this  problem.  It  was  soon  evident  that 
the  stability  was  dependent  upon  a  number  of  factors.  Among  these 
was  the  vitamine  content  of  the  starting  material  used,  the  chemical 
reaction  of  the  substrate  and  the  duration  of  the  heating  period.4 
Although  these  points  are  of  great  practical  importance  as  regards 
the  etiologic  and  dietetic  significance,  they  are  only  of  moderate 
theoretical  interest,  so  long  as  the  vitamine  itself  has  not  been  pre- 
pared in  the  pure  state  and  its  characteristics  determined.  Only 
then  shall  we  be  in  a  position  to  conduct  such  experiments  exactly. 
From  the  description  of  the  isolation  experiments,  it  is  clear  that  this 
vitamine  is  quite  stable  and  can  withstand  a  number  of  chemical  and 
physical  manipulations.  Therefore,  it  is  hard  to  determine  at  which 
stage  of  the  work  this  substance  loses  its  activity.  In  the  fraction- 
ation,  there  is  very  little  loss  of  activity  till  we  come  to  the  last 
stages,  where  it  decreases  rapidly.  One  almost  has  the  impression 
that  the  substances  accompanying  the  vitamine  serve  to  stabilize  it. 
We  must  admit  that  we  are  still  in  the  dark  as  to  the  cause,  and 
although  this  may  eventually  find  quite  a  simple  explanation,  it 
seems  to  us  for  the  present  remarkable  and  mysterious. 

As  regards  the  solubility  of  vitamine  B,  this  has  already  been 
spoken  of  in  the  preceding  pages.  All  investigators  apparently  agree 
that  this  substance  is  insoluble  in  strong  alcohol.  There  are  also 
some  experiments  which  indicate  that  prolonged  boiling  in  alcoholic 
solution  may  inactivate  this  vitamine.  As  to  the  effect  of  heat,  the 
findings  vary ;  similarly,  with  the  effect  of  alkalis.  On  the  other  hand, 
the  investigators  are  one  in  stating  that  this  vitamine  is  very  resistant 
to  acid.  Among  other  chemical  reagents,  diazotization,  according  to 

*Emmett  and  Luros  (I.e.  94)  give  a  good  bibliography  relative  to  this 
question. 


THE   ANTIBERIBERI   VITAMINE  195 

McCollum  and  Simmonds  (I.e.  315),  does  not  affect  the  activity  of 
vitamine  B,  and  this  has  recently  been  confirmed  by  Funk  and  Dubin 
(I.e.  493)  who  also  showed  that  the  substance  stimulating  the  growth 
of  yeast  was  not  destroyed  by  reduction  with  palladium  and  hydrogen, 
as  well  as  with  zinc  and  hydrochloric  acid ;  neither  was  it  affected  by 
oxidation  with  finely  divided  platinum  and  oxygen  for  7  to  8  hours. 
Besides  this,  we  (512)  showed  in  1916  that  the  action  of  radium  in 
doses  used  in  radium  therapy  is  without  destructive  influence  on  vita- 
mine  B  originating  from  autolyzed  yeast.  In  accord  with  this,  there 
is  the  work  of  Zilva  (513)  who  investigated  the  effect  of  ultra-violet 
light  on  this  substance.  Contrary  to  the  above  findings,  Sugiura 
and  Benedict  (514)  report  that  large  doses  of  X-rays  of  radium  almost 
obliterates  the  vitamine  of  autolyzed  as  well  as  dried  yeast.  Weill 
and  Mouriquand  (515)  carried  out  analogous  experiments  with 
Rontgen  rays  on  barley  kernels. 

DEMONSTRATION   AND   ESTIMATION   OF   ANTIBERIBERI   VITAMINE 

At  the  outset  of  vitamine  investigations,  only  one  method  was 
available  for  the  demonstration  of  vitamine  B.  Beriberi  was  pro- 
duced in  chickens  and  pigeons  (the  latter  being  more  commonly 
used)  by  feeding  white  rice,  and  then  the  preparation  to  be  tested 
was  given  either  per  os  or  intramuscularly.5  The  latter  method  had 
the  advantage  of  giving  a  more  speedy  effect  and  of  making  it  certain 
that  the  solution  was  utilized  by  the  animal,  and  had  not  trickled 
out  of  the  beak.  A  negative  finding  of  this  therapeutic  measure 
did  not  entirely  indicate  that  the  solution  to  be  tested  was  inactive. 
Negative  results  may  be  due  to  the  fact  that  the  animal  had  pro- 
gressed too  far  to  be  affected  by  the  above  measures  or  they  might 
be  due  to  the  presence  of  toxic  products.  It  was  chiefly  through  the 
efforts  of  Osborne  and  Mendel,  as  well  as  McCollum  and  his  co- 
workers,  that  the  second  method  was  then  developed,  of  which  we 
have  already  spoken  in  the  chapter  on  the  vitamine  requirements  of 
rats.  This  method  consists  in  adding  the  preparation  to  be  tested 
to  a  diet  complete  in  everything  but  vitamine  B.  If  growth  was 
obtained  with  young  rats,  then  the  presence  of  this  vitamine  was 

5  We  were  one  of  the  first  to  make  use  of  the  parenteral  administration  of 
this  vitamine.  Unfortunately,  we  have  been  unable  to  discover  in  the  liter- 
ature by  whom  this  method  was  first  introduced. 


196  THE   VITAMINES 

demonstrated.  This  method  is  more  certain,  though  it  has  the  draw- 
back that  much  more  time  is  required  for  its  application.  Both  of 
these  methods  have  the  great  disadvantage  that  in  isolation  experi- 
ments, every  new  fraction  must  await  the  findings  of  the  animal 
experiments,  the  vitamine  preparation  in  the  meantime  being  subject 
to  more  and  more  decomposition.  For  these  reasons,  efforts  have 
long  been  made  to  develop  a  method  that  would  permit  of  conducting 
a  test  in  the  shortest  possible  time.  For  practical  reasons  too,  such 
a  method  would  be  of  advantage  in  determining  the  vitamine- content 
of  various  foodstuffs. 

The  first  step  in  this  direction  was  made  by  Fraser  and  Stanton 
(I.e.  55) .  Based  upon  Schaumann's  theory  of  the  lack  of  phosphorus, 
they  believed  that  the  vitamine  content  of  rice  could  be  estimated 
by  the  amount  of  phosphorus  present.  Voegtlin  and  Myers  (516) 
also  suggested  the  phosphorus  content  of  wheat  and  corn  as  a  some- 
what reliable  indication  of  the  amount  of  vitamine  contained  therein. 
On  the  other  hand,  Green  (517)  showed  that  if  the  phosphorus 
content  of  American  corn  is  taken  as  a  unit,  then  all  South  African 
varieties  of  corn,  which  are  very  poor  in  phosphorus,  might  be 
regarded  also  as  very  poor  in  vitamines,  which  is,  however,  not  the 
case.  Ottow  (518)  is  likewise  of  the  opinion  that  the  estimation  of 
the  phosphorus  content  alone  may  lead  to  false  conclusions.  He 
believed  that  the  determination  of  the  quantity  of  alcohol-soluble 
fraction  of  rice  was  more  reliable,  at  the  same  time  regarding  the  use 
of  animal  experiments  as  the  only  method  to  be  relied  upon. 

We  (519)  sought,  at  first,  to  form  some  conception  as  to  the 
amount  of  vitamine  present,  by  determining  the  nitrogen  in  foodstuffs 
having  a  slight  non-protein  nitrogen,  such  as  milk.  For  this  purpose, 
we  dried  some  milk,  made  an  alcoholic  extract  of  the  portion  precipit- 
able  by  phosphotungstic  acid,  and  analyzed  it  for  its  nitrogen  content. 
The  only  result  apparent  was  that  on  centrifuging  the  milk,  a 
large  part  of  the  residual  nitrogen  is  lost  and  evidently  goes  over 
into  the  cream.  Whereas  in  a  non-centrifuged  milk  this  fraction 
amounts  to  about  2.2  mgm.  per  liter,  in  the  centrifuged  milk  it  is 
only  1.4  mgm.  This  shows  perhaps  that  milk  fat  (butter)  may 
contain  nitrogenous  vitamine.  Furthermore,  since  we  noted  that 
vitamine  fractions  contained  substances  whose  nitrogen  could  be  only 
partially  estimated  by  means  of  a  Kjeldahl  determination,  we  wanted 
to  make  use  of  the  difference  between  the  analyses,  according  to 


THE   ANTIBERIBERI   VITAMINE  197 

Dumas  and  to  Kjeldahl,  as  an  indication  of  the  vitamine  fraction. 
However,  since  this  fraction  contained  a  relatively  large  amount  of 
pyridine  derivatives,  the  method  is  valueless  so  long  as  we  do  not 
know  the  relationship  between  these  substances  and  the  vitamines. 
Brill  and  Alincastre  (520)  tried,  in  this  way,  to  average  the  maximum 
vitamine  content  from  the  phosphotungstic  acid  precipitates  of 
various  vegetables.  Seidell  (521)  sought  to  determine  the  vitamine 
content  of  yeast  by  estimating  the  nitrogen  in  activated  fuller's 
earth.  Eddy  (522)  tried  to  do  the  same  with  an  activated  fuller's 
eaiih  out  of  pancreas.  Since  fuller's  earth  adsorbs  also  a  large 
amount  of  inactive  material,  this  method  is  of  no  special  consequence. 
In  discussing  the  chemistry  of  vitamine  B,  it  will  be  recalled  that 
we  have  often  made  use  of  a  reaction  introduced  by  Folin  and 
Macallum  (I.e.  486)  for  the  determination  of  uric  acid  and  phenol. 
It  was  shown  by  us,  and  later  by  others,  that  all  vitamine-containing 
extracts  give  this  reaction.  It  depends  upon  the  development  of  a 
blue  color  with  phosphotungstic  and  phosphomolybdic  acids  on  the 
addition  of  sodium  carbonate.  The  chemical  nature  of  the  sub- 
stances giving  this  reaction  was  investigated  by  Funk  and  Macallum 
(523),  as  well  as  by  Lewis  and  Nicolet  (524).  With  Macallum,  we 
also  showed  that  if  an  incision  is  made  in  the  maize  kernel  and  the 
test  applied  in  situ  the  blue  color  develops  in  the  part  known  to  be 
rich  in  vitamine.  Naturally,  these  parts  are  even  richer  in  other 
extractives.  As  this  method  came  into  use  for  the  chemical  analysis 
of  various  corn  meal  products,  we  (525)  showed  that  in  extracts, 
prepared  either  cold  or  warm,  the  developed  color  increased  with  the 
vitamine  content.  In  the  warm  extracts,  the  color  was  weaker, 
showing  that  the  substances  responsible  for  the  color  reaction  are 
destroyed  by  hfe'at.  If  it  should  appear  later,  with  greater  certainty, 
that  this  reaction  has  nothing  in  common  with  the  vitamine,  it  might 
still  be  useful  in  vitamine  fractionation  as  an  index  of  the  purity  of 
the  isolated  substances.  These  reactions  could  conceivably  also 
serve  as  an  index  of  the  purity  of  the  isolated  lipoids. 

Green  (526)  tried  to  estimate  the  vitamine  content  of  various  food- 
stuffs by  means  of  a  biological  method  (animal  experiments).  The 
method  depends  on  the  length  of  time  pigeons  survive  on  various 
diets,  rich-  or  poor  in  vitamines.  As  a  result  of  these  studies,  Green 
proposed  the  following  formula: 

S  =  V  -  X  '  K 


198  THE    VITAMINES 

in  which  S  is  the  duration  of  life;  C,  the  amount  of  vitamine  that 
the  animal  may  lose  without  developing  beriberi;  F,  the  amount  of 
vitamine  necessary  to  provide  for  the  metabolism  of  a  given  quantity 
of  food;  X,  the  quantity  of  vitamine  in  the  given  diet;  K,  the  con- 
stant depending  upon  the  quality  and  quantity  of  the  diet  fed.  By 
means  of  this  formula,  the  vitamine  content  of  various  foodstuffs, 
especially  corn,  was  determined.  The  figures  obtained  correspond  in 
general  with  experience  gained  by  other  investigators  with  the  same 
products.  The  weak  point  of  the  formula  is  that  it  shows  the  pres- 
ence of  vitamine  in  white  rice  despite  all  evidence  to  the  contrary. 
Aside  from  this,  it  is  obvious  that  the  survival  of  pigeons  depends  on 
many  unknown  factors. 

We  come  now  to  the  consideration  of  a  method,  briefly  discussed 
in  the  chapter  on  the  vitamine  requirements  of  yeast,  which  was 
indicative  of  real  progress.  This  method  is  b'ased  upon  the  fact 
that  certain  yeasts  require  an  addition  of  substances  containing 
vitamines  in  order  to  grow.  We  have  already  spoken  of  the  work  of 
Williams  (I.e.  128),  Bachmann  (I.e.  129)  and  Abderhalden  and 
Schaumann  (I.e.  499) — all  of  which  led  to  the  development  of  this 
method.  Soon  after,  papers  were  published  on  the  practical  applica- 
tion of  the  method.  Eddy  and  Stevenson  (527)  tested  the  method 
of  Bachmann  and  found  that  it  was  not  so  very  reliable.  Thereupon 
they  turned  to  the  method  of  Williams,  by  introducing  exactly 
calibrated  micropipettes  (similar  to  those  used  in  opsonic  index 
determinations)  for  drawing  up  the  yeast  cell  suspension  and  the 
vitamine  solution,  which  made  the  method  somewhat  more  exact. 
The  yeast  and  vitamine  units,  with  the  necessary  controls,  were 
deposited  on  a  counting  slide  and  kept  in  the  incubator.  After  a 
certain  time,  the  slides  were  removed  from  the  incubator,  stained 
and  the  cells  counted.  The  objection  to  this  method  is  the  fact 
that  clumps  of  yeast  cells  are  often  introduced  with  the  cell  suspen- 
sion. In  spite  of  shaking  the  yeast  cell  suspension  and  partial 
centrifugation  to  remove  the  clumps,  this  objection  was  not  quite 
overcome  and  the  results  were  therefore  uncertain.  Among  other 
substances,  Eddy  and  Stevenson  also  tested  the  products  isolated  by 
us  from  yeast  in  1912  and  1913  (see  chapter  on  the  Chemistry  of 
Yeast)  and  found  them  active,  while  nicotinic  acid  was  inactive.  The 
active  substance  was  not  destroyed  by  heating  to  100°C.,  but  partially 
so  at  120°C.  Using  this  method,  more  vitamine  was  found  in  the 
mammary  vein  than  in  the  jugular. 


THE    ANTIBERIBERI   VITAMINE 


199 


The  methods  of  R.  J.  Williams  (528)  and  of  Funk  and  Dubin  (I.e. 
511)  then  appeared  almost  simultaneously,  both  signifying  a  step 
forward  in  the  demonstration  of  the  part  played  by  the  vitamines 
in  yeast  growth.  The  method  of  Williams  is  as  follows :  0.3  gram  of 
ordinary  fresh  bakers'  yeast  was  removed  from  the  center  of  a  yeast 
cake  and  suspended  in  a  liter  of  sterile  water.  Of  this  suspension, 
1  cc.  was  introduced  into  the  nutritive  solution  and  the  cultures  kept 
in  the  incubator  at  30°C.  for  eighteen  hours,  after  which  growth  was 
stopped  by  the  addition  of  formalin.  The  yeast  cells  were  then 


12 


E    |0 

0) 

f    6 


3 

O 

to 
OS 


•*> 


I  a..        3  4  .5 

Vltamine  solution— cc. 

FIG.  47.  INFLUENCE  OP  VITAMINE  B  CONCENTRATION  ON  YEAST  GROWTH 

(WILLIAMS) 

filtered  on  a  Gooch  crucible,  washed  with  water  and  alcohol  and 
dried  at  103°C.  for  2  hours.  With  a  vitamine  solution  of  a  source 
not  mentioned,  a  growth  curve  was  obtained,  reproduced  herewith. 
He  showed  that  with  higher  vitamine  concentrations,  the  quantity 
of  cells  rose  proportionally  and  that  under  certain  conditions,  the 
curve  approaches  a  straight  line.  Williams  also  observed  that  certain 
extracts  such  as  autolyzed  yeast,  or  extracts  heated  to  a  high  tem- 
perature in  the  presence  of  acids,  could  exert  an  inhibiting  influence 
on  the  growth  of  yeast.  In  such  cases,  an  abundant  dilution  is  of 
great  help  and  yields  better  results. 


200 


THE    VITAMINES 


The  method  of  Funk  and  Dubin  is  still  more  simple  and  is  briefly 
as  follows:  A  yeast  suspension  is  prepared  by  shaking  100  cc.  of 
Nageli  solution  inoculated  with  a  platinum  loop-full  of  a  48  hours 
pure  yeast  culture,  preferably  bottom  yeast.  Two  test  tubes  are 


FIG.  48.  CENTRIFUGE  TUBES  USED  IN  THE  DETERMINATION  OF  THE  EFFECT 
OF  VlTAMINE  ON  YEAST  GROWTH 

The  tube  at  the  left  shows  the  reading  of  the  blank,  while  the  other  two 
tubes  show  the  readings  obtained  with  different  amounts  of  autolyzed  yeast 
(Funk-Dubin). 

prepared,  containing  the  unknown  vitamine  solution  to  be  tested; 
in  addition,  there  are  two  controls — one  with  yeast  suspension  alone, 
and  one  with  the  vitamine  solution  alone.  The  last  control  is  not 
very  important  and  serves  only  to  detect  the  formation  of  a  pre- 


THE    ANTIBERIBEKI   VITAMINE 


201 


cipitate  or  cloudiness  at  the  beginning  of  the  test.  All  of  the  test 
tubes  are  placed  in  the  incubator  for  20  hours  at  30°C.,  after  which 
the  cells  are  inactivated  by  immersing  the  tubes  in  a  water  bath  at 
70  to  80°C.  for  a  few  minutes.  The  contents  of  the  tubes  are  then 
quantitatively  transferred  to  centrifuge  tubes,  the  lower  part  of 
which  is  in  the  form  of  a  capillary,  graduated  in  mm.  The  tubes 
are  centrifuged  for  15  minutes  at  about  2600  r.p.m.,  and  the  volume 
of  cells  read  on  the  graduated  capillary  of  the  centrifuge  tube.  The 
reading  should  be  taken  at  once,  since  there  is  a  tendency  for  the 
yeast  cells  to  swell  up,  after  a  time.  The  control,  without  the 
addition  of  vitamine,  must  be  subtracted  from  the  reading. 

We  found  that  the  controls  with  yeast  alone  were  quite  constant 
and  almost  independent  of  the  number  of  yeast  cells  in  the  suspension 


Mm. 
5 


0.5        1.0 


1.5 


Z.O        2.5        aO        3.5       4.0        4.5       5.0 
Cc  of  yeast  suspension 

TIG.  49.  EFFECT  OF  VARYING  AMOUNTS  OF  YEAST  SUSPENSION  (FUNK-DUBIN) 

used,  as  may  be  seen  from  the  curve.  Experiments  with  increasing 
amounts  of  autolyzed  yeast  gave  a  curve  markedly  different  from 
that  of  Williams,  but  which  is  perhaps  better  applicable  to  the  study 
of  natural  extracts.  We  feel  that  our  results  have  been  considerably 
influenced  by  the  presence  of  inhibiting  substances.  For  this  and 
other  reasons,  it  appears  somewhat  premature  to  estimate  the 
vitamine  content  of  various  foodstuffs  by  the  use  of  these  methods. 
A  number  of  problems  were  undertaken  with  the  help  of  the  above 
method,  and  we  shall  record  some  of  the  results.  We  showed  that 
analyzed  substances,  isolated  from  yeast  seven  years  ago  by  the 
author,  still  retained  a  definite  activity,  although  they  had  been 
re  crystallized  to  constant  melting  point.  On  the  other  hand,  nico- 
tinic  acid  obtained  from  yeast  was  inactive.  White  polished  rice 


202 


THE   VITAMINES 


showed  no  influence  on  yeast  growth;  this  was  also  true  of  vitamines 
A  and  C  (after  vitamine  B  had  been  completely  removed  by  adsorp- 
tion with  fuller's  earth).  The  active  substance  was  found  also  out- 
side of  the  wheat  and  corn  germ,  though  in  smaller  amounts.  The 
substance  influencing  the  growth  of  yeast  was  very  resistant  towards 
heat;  it  could  be  diazotized,  oxidized  and  reduced  without  loss  of 
activity.  With  this  method  as  a  control,  a  fractionation  of  yeast 
was  undertaken;  partial  results  have  already  been  published  (Funk 
and  Dubin  (I.e.  511)  )  but  the  work  is  not  yet  completed. 

Mm. 
35 


30 
25 
20 
15 
10 


7 


0.8 


0.9        i.O 


0.0        0.1         0.2        0.3        0.4        0.5        0.6        0.7 
Cc.  of  autolyzed  yeast 

FIG.  50.  VITAMINE  ACTIVITY  SHOWN  BY  INCREASING  AMOUNTS  OF  AUTOLYZED 
YEAST  (FUNK-DUBIN) 

The  use  of  these  methods  was  criticized  by  de  Souza  and  McCollum 
(529).  They  employed  the  first  method  of  Williams,  slightly 
modified,  for  their  investigations  and  found,  as  a  rule,  that  with 
increasing  amounts  of  vitamine  B,  the  growth  of  yeast  is  favorably 
affected.  They  believed,  however,  that  the  method  could  hardly  be 
applied  to  the  determination  of  vitamine  B,  because  glucose  and 
ammo-acids  also  show  a  growth-promoting  action.  Despite  this,  if 
we  examine  their  findings  more  closely  we  see  that  the  addition  of  a 
10  per  cent  glucose  solution  had  no  more  influence  during  the  first 
24  hours,  as  compared  with  the  controls;  it  was  only  after  48  hours— 


THE   ANTIBERIBERI   VITAMINE  203 

• 

and  not  always  so — that  some  effect  was  noticeable.  This  protrac- 
tion of  the  influence  showed  that  the  accelerative  action  of  glucose 
was  of  a  secondary  nature,  and  might  perhaps  be  explained  by  the 
plasmolysis  of  some  yeast  cells.  When  de  Souza  and  McCollum  speak 
of  the  favorable  action  of  amino  acids,  they  mean  by  that  a  product 
resulting  from  the  sulphuric  acid  hydrolysis  of  meat.  This  prepara- 
tion, as  well  as  an  alcoholic  extract  of  wheat  germ,  was  shown  to  be 
free  from  vitamine  B  in  rat  experiments,  while  they  were  still  active 
for  yeast.  Hydrolyzed  meat  gave  only  double  the  amount  of  cells, 
so  that  the  action  was  not  so  pronounced.  These  investigators 
admitted  that  their  experiment  did  not  demonstrate  that  vitamine  B 
played  no  part  in  the  growth  of  yeast,  they  likewise  showed  that  the 
growth  of  rats  and  yeast  does  not  go  parallel  with  the  same  extracts. 
The  question  as  to  the  identity  of  vitamine  B  with  the  substance, 
promoting  the  growth  of  yeast  will  be  discussed  in  the  next  section. 
Newer  investigations  have  shown  that  the  yeast  test  is  of  value 
in  determining  the  relative  richness  of  extracts  in  water-soluble 
vitamines,  but  cannot  be  used  for  the  quantitative  determination 
of  vitamine  B. 

THE    POSSIBLE    IDENTITY    OF   VITAMINE    B    WITH    THE    SUBSTANCE 
STIMULATING   THE   GROWTH   OF  ANIMALS   AND   OF   YEAST 

We  have  already  (p.  41)  touched  upon  the  question  raised  by 
Mitchell  (I.e.  93)  as  well  as  by  Emmett  and  Luros  (I.e.  94),  as  to 
the  identity  of  these  substances.  As  regards  the  comparison  of 
vitamine  B  with  the  substance  promoting  the  growth  of  rats,  Funk 
and  Macallum  (530)  showed  that  the  latter  substance  could  be 
precipitated  with  phosphotungstic  acid,  whereupon  further  fraction- 
ation  was  tried.  Since  the  diet  we  used  at  the  time  did  not  contain 
sufficient  vitamine  A,  the  results  were  not  particularly  fruitful,  and 
seemed  to  indicate  that  pigeons  require  less  vitamine  B  for  protection 
against  beriberi  than  do  rats  for  growth,  or  that  vitamine  B  is  more 
stable  than  the  growth  vitamine.  Drummond  (I.e.  498)  corrob- 
orated our  findings  and  came  to  the  conclusion  that,  in  all  prob- 
ability, both  substances  are  identical.  Emmett  and  Luros  (I.e.  94) 
seem  to  have  a  different  opinion  on  this  matter.  They  investigated 
the  effect  of  autoclaving  at  various  temperatures  for  various  periods 
of  time.  On  the  one  hand,  these  products  were  given,  as  such,  to 


204  THE   VITAMINES 

I 

pigeons  and  on  the  other,  together  with  lactalbumin,  butter,  salts  and 
lard,  to  rats.  Vitamine  B,  which  is  necessary  for  pigeons,  appeared  to 
be  unstable  and  was  totally  destroyed  by  heating  for  2  hours  at 
1 20°C . ,  while  rats  could  still  grow  on  the  above  food  mixture .  Emmett 
and  Luros  came  to  the  conclusion  that  these  vitamines  are  different 
from  each  other.  Nevertheless,  we  saw  in  our  investigations  on 
yeast,  contrary  to  the  above  experiment,  that  vitamine  B  was  resis- 
tant to  heat ;  furthermore,  in  the  experiments  of  these  workers  one  kind 
of  animal  was  fed  rice  alone  while  the  other  received  various  addi- 
tions. As  we  shall  see  later,  protein  may  easily  take  the  place  of  a 
vitamine  addition,  besides  which,  these  animals  may  differ  with 
respect  to  their  vitamine  requirements. 

As  regards  the  identity  of  the  growth  substance  for  yeast  with 
vitamine  B,  the  question  is  of  particular  importance  since  we  must 
know,  when  using  the  yeast  method  described  in  the  previous  chapter, 
which  substance  we  are  determining.  There  is  no  doubt,  however, 
that  the  substance  important  for  the  growth  of  yeast  is  much  more 
stable  to  heat,  and  yet  we  saw  in  the  chapter  on  the  chemistry  of 
vitamines,  that  vitamine  B  resists  many  manipulations,  and  still 
retains  its  activity  after  many  weeks  of  work.  Emmett  and  Stock- 
holm (531),  on  the  contrary,  saw  that  strongly  heated  unpolished 
rice,  although  inactive  for  pigeons,  is  just  as  active  as  the  natural 
product  for  yeast  cells.  The  same  findings  were  obtained  with  ni- 
trates resulting  from  the  treatment  of  extracts  with  fuller's  earth. 
In  these  cases,  the  filtrates  were  active  for  yeast  but  not  for  rats.6 
Emmett  and  Stockholm  believed  that  further  research  must  establish 
either  the  identity  or  dissimilarity  of  these  substances,  and  it  would 
not  be  correct  in  the  meanwhile,  to  regard  the  yeast  method  as 
suitable  for  the  determination  of  vitamine  B. 

Recently,  Funk  and  Dubin  (I.e.  129d)  have  corroborated  the 
statement  of  Emmett  and  Luros  that  the  yeast  substance  is  not 
identical  with  vitamine  B.  If  autolyzed  yeast  is  shaken  with  a 
small  amount  of  fuller's  earth  vitamine  B  is  usually  removed  almost 
quantitatively.  Now  if  the  filtrate  from  the  above  is  repeatedly 
shaken  with  larger  amounts  of  fuller's  earth,  the  substance  acting 

6Karr  (I.e.  404),  working  on  dogs,  noted  that  vitamine  B  of  yeast  is  very 
stable.  Autoclaving  for  3  to  4  hours  with  subsequent  heating  at  108°C.  for 
72  hours  destroyed  only  a  part  of  this  substance. 


THE   ANTIBERIBERI   VITAMINE  205 

on  yeast — and  which  we  call  vitamine  D — is  likewise  almost  com- 
pletely removed.  We  have  not  succeeded  thus  far  in  separating 
D-vitamine  from  B-vitamine  quantitatively.  The  animal  experi- 
ments performed  thus  far  in  which  vitamine  B  preparations  were 
used  have  to  be  repeated,  since  such  preparations  contain  at  least 
two  vitamines — B  and  D.  While  pigeons  get  along  very  well  on 
vitamine  B  alone,  rats  require  vitamine  D  besides  vitamine  B. 
Other  animals  are  being  investigated  in  this  respect. 

Vitamine  D  is  one  of  the  substances  necessary  for  the  growth  of 
certain  bacterias,  as  we  have  already  pointed  out.  The  isolation 
has  been  attempted  by  Goy  (531a)  who  thought  it  was  a  non- 
nitrogenous  substance  but  the  pure  substance  he  isolated  proved 
to  be  inactive.  The  isolation  also  was  attempted  by  Frankel  and 
Schwartz  (531b). 

We  also  feel  that  this  subject  needs  further  investigation.  For 
in  every  one  of  the  cases  investigated,  we  tested  the  activity  after 
the  naturally  occurring  substances  had  been  altered  by  the  manipu- 
lation undergone;  we  do  not  know  what  the  action  of  the  substances 
in  the  natural  state  would  have  been.  It  may,  for  example,  be 
established  later  that  vitamine  B  is  a  uniform  substance,  but  that 
parts  of  it  may  be  utilized  differently  by  yeast,  pigeons  and  rats. 
The  analogy  between  the  three  modes  of  action,  in  relation  to  its 
occurrence  in  Nature,  is  so  far  extended  that  we  are  perhaps  justi- 
fied in  attributing  to  it  a  genetic  relationship.  The  findings  of 
Funk  and  Dubin  (I.e.  493),  and  others,  as  to  specificity,  also  point 
to  this  conception. 

SPECIFICITY   OF   ANTIBERIBERI   VITAMINE 

A  short  time  after  the  appearance  of  our  chemical  vitamine  studies, 
a  number  of  publications  appeared  describing  the  curative  action  on 
beriberi  of  various  quite  heterogeneous  substances.  These  papers 
had  the  regrettable  effect  that  doubt  arose  as  to  the  specificity  of 
vitamine  influence.  It  almost  seemed  as  though  practically  all 
chemical  groups,  under  certain  conditions,  could  act  as  vitamine. 
Although  most  of  these  results  were  either  disproven  or  else  aban- 
doned by  their  respective  authors,  we  see  unfortunately  that  these 
results  gained  acceptance  in  the  literature.  We  must  not  forget 
that  the  cure  of  acute  beriberi  symptoms  may  be  accomplished 
theoretically  in  two  ways,  first,  by  the  addition  of  the  specific  vita- 


206  THE    VITAMINES 

mine,  second,  by  measures  which  will  suddenly  mobilize  the  reserve 
vitamiiie  present  in  organs  and  tissues.  As  regards  spontaneous 
cures,  which  we  personally  have  not  seen  (but  which  seem  to  be 
plausible),  occurring  ostensibly  in  starvation,  we  may  explain  them 
by  the  catabolism  of  the  tissues  and  the  consequent  liberation  of  the 
vitamine.  Every  substance,  having  pharmacological  action,  which 
stimulates  metabolism  may  also  contribute  to  the  above-mentioned 
action. 

Eijkman  (532)  stated  that  pigeons,  used  by  us  for  the  demonstra- 
tion of  vitamine  B,  were  not  suitable  for  this  purpose,  since  they 
show  a  different  behavior  from  chickens.  He  maintained  that  a 
mixture  of  potassium  chloride  and  sodium  chloride  (3:1)  in  doses  of 
20  to  40  mgm.  can  cure  beriberi  in  pigeons,  but  not  in  chickens.  We 
(533)  tested  this  out  on  pigeons,  but  could  not  corroborate  Eijk- 
man. In  spite  of  this,  we  note  that  his  findings  are  frequently 
quoted  in  the  literature.  Later,  we  showed  (534)  that  certain  purine 
and  pyrimidine  derivatives  exerted  a  life-prolonging  action  on  pigeons 
fed  on  rice.  This  observation  was  confirmed  by  some,  Williams  and 
Saleeby  (I.e.  472),  and  refuted  by  others.  Since  these  products  are 
frequently  not  indifferent  pharmacologically,  their  behavior,  as 
described  above,  may  perhaps  be  explained  in  that  light.  Cooper 
(535  and  536)  found  that  quinine  and  strychnine  exhibited  a  definite 
action  on  experimental  beriberi.  Later,  he  showed  that  quinine 
contained  vitamine  as  an  impurity;  as  for  the  action  of  strychnine 
it  may  be  explained  by  its  effect  on  the  metabolism.  Abderhalden 
and  Lampe  (I.e.  25)  stated  that  castor  oil  had  a  curative  effect,  but 
later  on  Abderhalden  and  Ewald  (537)  reported  that  they  could  not 
confirm  the  earlier  finding.  They  believed,  however,  that  vitamine 
B  was  analogous  to  B-imidazolethylamine.  Dutcher  (I.e.  .276) 
investigated  the  behavior  of  thyroid,  thyroxin  (chemically  pure 
active  substance  of  thyroid),  tethelin  (lipoid  from  the  hypophysis) 
and  pilocarpine  and  found  that  these  substances  have  a  therapeutic 
effect  on  beriberi.  Pilocarpine  was  found  inactive  by  Abderhalden 
and  Ewald;  thyroid  was  regarded  as  inactive  by  Vedder  and  Clark 
(I.e.  266).  Seaman  (537a),  on  the  contrary,  could  demonstrate  the 
presence  of  vitamine  B  in  thyroid.  Inasmuch  as  thyroid  substance 
acts  as  a  powerful  stimulus  to  the  metabolism,  the  mobilization  of 
the  vitamine  from  the  tissues  may,  under  certain  circumstances, 
simulate  a  curative  action,  although  we  convinced  ourselves  that 


THE    ANTIBERIBERI    VITAMINE  207 

death  is  hastened  because  of  it.  Regarding  the  action  of  tethelin, 
when  one  considers  its  method  of  preparation,  it  may  contain  vita- 
mine  as  an  impurity.  Butcher,  Holm  and  Bierman  (511)  tested 
various  indol  derivatives  of  thyroxin  and  found  them  active,  but 
their  findings  require  further  corroboration.  To  complete  our  data 
on  this  question,  we  may  mention  that  it  was  impossible  for  Chamber- 
lain, Vedder  and  Williams  (I.e.  64)  and  Abderhalden  and  Schaumann 
(I.e.  499)  to  influence  beriberi  by  the  administration  of  known  sub- 
stances, and  especially  a  mixture  of  amino  acids.  Viewing  all  state- 
ments to  the  contrary  in  a  critical  light,  we  must  recognize  that 
vitamine  B  has  a  well  denned  and  specific  action,  and  it  is  not  to  be 
substituted  by  other  products. 

PHYSIOLOGY  AND  PHARMACOLOGY  OF  ANTIBERIBERI  VITAMINE 

To  draw  a  general  picture  of  the  nature  of  beriberi,  we  see,  first  of 
all,  a  very  definite  retardation  of  the  life  processes,7  similar  to  that 
observed  in  inanition.  In  this  respect,  Ramoino  (538)  showed  that 
in  rice-fed  pigeon,  the  respiratory  quotient  is  very  low  but  may  be 
raised  again  by  an  addition  of  vitamine.  Similar  observations  were 
made  by  Jansen  and  Mangkoewinoto  (539).  This  is  evident  also 
in  McCarrison's  (I.e.  300)  investigations  of  avian  beriberi,  in  which 
he  described  pathological  changes  in  the  intestines  and  some  glandular 
organs.  In  hunger,  however,  the  whole  organism  is  attuned  to 
endure  the  least  possible  loss  of  energy  and  substance.  On  the 
other  hand,  in  experimental  beriberi  only  a  diet  free  from  or  poor  in 
vitamines  is  fed.  In  this  way,  energy  is  expended,  digestive  juices  are 
secreted  and  the  assimilative  processes  are  continued.  At  present,  we 
are  not  quite  in  a  position  to  set  forth  the  primary  causes  of  the  whole 
disturbance.  It  is  not  yet  possible  to  say,  of  the  pathological  con- 
ditions observed  in  this  disease,  which  may  be  regarded  as  primary 
and  which  secondary.  We  only  know  that  the  animal  requires  a 
minimal  amount  of  vitamine  in  order  to  regain  its  normal  condition. 
With  one  stroke,  the  entire  condition  is  altered,  and  it  can  hardly 
bo,  believed  that  this  is  brought  about  by  such  a  small  quantity  of 

7  In  accord  with  this,  there  are  the  observations  of  Drummond  (I.e.  33 
and  Abderhalden  (I.e.  500)  on  the  subnormal  temperatures  of  beriberi 
pigeons. 


208 


THE    VITAMINES 


substance.8  In  order  to  find  an  analogy,  we  may  compare  an  animal 
on  a  vitamine  free  diet  with  a  machine,  running  without  oil.  At 
first  the  machine  operates  faultlessly,  but  gradually  trouble  arises 
and  then,  in  due  time,  greater  defects  occur  which  can  nevertheless 
still  be  corrected  by  proper  oiling.  In  the  end,  a  condition  results 
which  can  no  longer  be  be  remedied  and  the  machine  falls  to  pieces. 
In  animals,  we  know  only  the  primary  cause,  lack  of  the  specific 
vitamine,  similar  to  the  oil  in  the  above  mentioned  analogy.  It 
still  seems  tempting  to  attribute  all  of  the  observed  phenomena  to 
changes  in  the  central  nervous  system,  but  another  explanation  is 
that  the  vitamine  might  be  thought  of  as  the  mother  substance  of  an 
important  product  of  the  endocrine  glands — a  conception  which  we, 
among  others,  thought  to  be  plausible  and  which  has  again  been 
put  forward  by  Massalongo  (540).  Another  relationship  may  be 
seen  in  the  origin  of  the  digestive  juices.  Lumiere  (541)  believed 
that  the  primary  cause  of  beriberi  is  to  be  noted  in  the  lack  of  secre- 
tions. The  same  view  was  held  by  Damianowich  and  Pilado  (541a). 
Other  investigators  sought  for  a  relationship  with  the  oxidation 
processes  in  the  body.  Butcher  (542)  and  Dutcher  and  Collatz 
(543)  demonstrated  a  decrease  in  the  catalase  in  the  organs  of  beriberi 
pigeons,  but  almost  the  same  thing  was  described  in  starving  animals 
by  Burge  and  Neill  (544).  Particularly  in  the  observations  as  to 
the  nature  of  beriberi,  is  the  picture  of  this  disease  somewhat  obscured 
by  complication  with  symptoms  of  inanition.  Jansen  (545)  studied 

8  The  influence  of  small  quantities  of  vitamines  on  the  metabolism  of 
pigeons  is  well  illustrated  in  the  metabolism  studies  of  Schaumann  (I.e.  403). 


VOLUNTARY  IN- 
GESTION  0  P 
WHITE  RICE 

CALORIES 

LOSS  OP  WEIGHT 

1 

t 

PERCENTAGE  RELATION  OF  OUTGO 
TO  INTAKE 

N 

P.O. 

Ash 

CaO 

MgO 

Without 
vitamine  
With  vitamine  .  .  . 

grams 

204.3 
295.1 

658.7 

951.6 

per  cent 

20.7 
9.41 

per  cent 

11.47 
8.53 

69.3 
101.4 

45.26 
63.56 

40.13 
62.12 

36.99 
118.9 

238.89 
347 

The  nitrogen  balance  was  positive  and  the  balance  of  the  inorganic  con- 
stituents was  likewise  improved.  The  weight  of  the  feces,  in  comparison 
with  the  food  intake,  showed  that  the  food  was  utilized  much  better. 


THE    ANTIBERIBERI   VITAMINE  209 

the  question  as  to  the  identity  of  secretin  with  vitamine  and  came 
to  the  conclusion  that  they  are  not  identical.  Voegtlin  and  Myers 
(546)  also  took  up  this  question  and  showed  that  both  substances 
occur  in  the  same  fraction;  they  believed  that  both  substances  at 
least  resemble  each  other,  if  they  are  not  entirely  identical.  The 
results  of  their  experiments  do  not  seem  to  lead  to  such  conclusions, 
since  they  noted  in  the  same  fraction  a  strong  secretin  and  a  weak 
vitamine  action,  whereas  a  vitamine  preparation  showed  a  consider- 
able vitamine  and  only  a  slight  secretin  content.  In  this  connection, 
they  showed  later  that  vitamine  fractions  were  contaminated  by  a 
histamine-like  substance.  That  the  substances  are  not  identical  was 
shown  by  the  work  of  Cowgill  (546a)  and  of  Anrep  and  Drummond 
(546b) .  Bickel,  Eisenhardt  and  Djenab  (547)  have  demonstrated  the 
presence  of  a  secretin  in  spinach,  which  stimulates  the  stomach  juice 
and  pancreas  secretion.  The  substance  occurred  in  the  arginine- 
histidine  fraction  and  was  inactivated  by  heating  to  140°C.  It 
occurred  in  combination  and  could  be  liberated  by  hydrochloric 
acid  hydrolysis.  The  relationship  of  this  substance  to  vitamines 
was  discussed  by  the  authors,  and  their  results  were  confirmed  by 
van  Eweyk  (548). 

Boruttau  (549)  believed  that  yeast,  or  the  aleurone  layer  of  oat 
kernels,  contains  a  substance  which  acts  as  a  specific  antidiabetic, 
since  their  extracts  greatly  decreased  the  output  of  sugar.  Uhlmann 
(550)  investigated  the  pharmacological  action  of  orypan  (extract  of 
rice  polishings),  as  well  as  the  extracts  of  various  vegetables  and  food- 
stuffs. The  action  noted  was  similar  to,  but  not  identical  with, 
that  of  choline,  and  pilocarpine.  Uhlmann  ascribed  this  to  the 
presence  of  vitamine.  It  is  nevertheless  apparent  that  in  his  cases, 
he  was  dealing  with  histamine-like  substances.  A  vitamine  prepara- 
tion made  by  us,  was  shown  by  Prof.  Cushny  to  have  no  action  on 
blood  pressure,  respiration  and  the  heart.  No  relationship  to  the 
endocrine  glands,  aside  from  that  already  described  in  avian  beriberi, 
was  noted. 

When  an  animal  is  fed  a  vitamine-free  food,  what  happens  to  the 
vitamine  reserves  stored  up  in  the  organs?  It  would  be  plausible  to 
assume  that  the  animal  would  hold  on  to  this  valuable  substance, 
but  this  is  not  the  case.  Cooper  (I.e.  536)  showed  that  when  feces 
of  chickens,  fed  on  unpolished  rice,  or  of  rabbits,  fed  on  bread  and 
cabbage,  are  extracted  with  alcohol  and  the  concentrated  extracts 


210  THE    VITAMINES 

given  to  beriberi  pigeons,  a  cure  could  be  obtained.  This  shows 
that  all  the  vitamine  of  the  food  is  not  assimilated,  but  that  a  portion 
of  it  appears  in  the  feces.  Another  explanation  might  be  found  in 
the  vitamine  content  of  the  bacterial  intestinal  flora,  although  Brad- 
don  and  Cooper  (551)  noted  only  a  small  quantity  of  vitamine  in 
these  bacteria.  Portier  and  Random  (I.e.  217)  believed  on  the  con- 
trary that  the  vitamine  of  the  intestinal  canal  arises  from  the  bacteria. 
Muckenfuss  (552)  found  vitamine  B  in  bile  and  urine,  and  was  con- 
firmed, as  regards  the  latter,  by  Gaglio  (553)  and  by  Funk  and  Dubin. 
Funk  showed  that  when  pigeons,  which  had  died  of  beriberi,  were 
minced,  extracted  with  alcohol,  the  extract  concentrated,  the  residue 
taken  up  with  water  and  given  to  other  beriberi  pigeons,  the  latter 
were  promptly  cured.  This  experiment,  corroborated  by  Theiler, 
Green  and  Viljoen  (I.e.  278)  showed  definitely  that  animals  develop 
beriberi  in  spite  of  the  fact  that  they  still  have  vitamine  in  their 
tissues.  It  seems  that  a  portion  of  the  vitamines  in  the  tissues  (per- 
haps as  a  constituent  of  the  cell  protoplasm)  is  so  combined  that  the 
animal  itself  cannot  utilize  it.  Another  explanation  of  this  is  to 
be  found,  perhaps,  in  the  work  of  Green  on  pigeons  (I.e.  526);  he 
noted  that  the  animal  organism  requires  a  rather  definite  amount  of 
vitamine  for  normal  health.  This  really  seems  to  be  the  case,  and 
as  the  quantity  drops  below  the  minimal  need,  it  leads  to  an 
avitaminosis.  The  quantity  of  vitamines  that  may  be  liberated  from 
the  organs  normally  is  very  small.  If  the  animal  is  given  a  diet  poor 
in  vitamine,  it  might  be  thought  that  the  animal  would  consume 
more  of  the  food  in  order  to  obtain  the  necessary  vitamine;  but  this 
is  not  the  case.  It  was  supposed  by  Osborne  and  Mendel  (554)  that 
some  foodstuffs  contain  sufficient  vitamine  for  rats,  if  these  animate 
would  but  ingest  more  of  this  food  mixture.  But  it  is  right  here 
that  we  meet  with  a  difficulty  that  is  also  of  practical  importance, 
namely,  the  question  of  vitamine  dilution  in  the  natural  foodstuffs. 
With  every  diet  prepared,  the  needed  amount  of  vitamine  must  be 
supplied  if  the  diet  is  to  prove  satisfactory. 

THE    INFLUENCE    OF   DIETAKY   COMPOSITION   ON   THE   VITAMINE 
REQUIREMENT 

Regarding  the  influence  of  a  large  or  small  addition  of  a  vitamine- 
free  diet  on  the  occurrence  of  beriberi,  the  literature  is  full  of  conflict- 
ing opinions.  While  Maurer  (555)  and  Cooper  (I.e.  536)  stated  that 


THE    ANTIBERIBERI    VITAMINS 


211 


the  addition  of  large  quantities  of  rice  hastens  the  onset  of  the 
disease,  Chamberlain,  Bloombergh  and  Kilbourne  (556)  noted  that 
the  disease  developed  sooner  on  a  smaller  rice  feeding.  They  found, 
in  fact,  that  beriberi  arises  in  hunger.  These  observations  were  later 
corroborated  by  Eijkman  and  Hoogenhuyze  (557) .  They  found  that 
the  outbreak  of  beriberi  could  be  obtained  by  starvation,  especially 
when  the  precaution  was  taken  to  wash  out  the  organism  with  water. 
The  starvation  beriberi  resulting  thereby  could  be  cured  on  the 
addition  of  vitamine.  At  all  events,  Theiler,  Green  and  Viljoen  (I.e. 
278)  noted  spontaneous  cures  in  animals  receiving  only  water.  These 
therapeutic  results  were  attributed  to  the  mobilization  of  the  vitamine 
present  in  the  tissues.  Walshe  (558)  found  no  beriberi  in  chickens 
that  had  been  starving  a  long  time;  these  observations  have  not  yet 
been  made  on  pigeons,  and  it  is  an  open  question  as  to  whether 
starvation  beriberi  may  develop  in  other  animals,  and  also  in  man. 
The  influence  of  the  composition  of  the  diet  on  the  occurrence  of 
beriberi  was  investigated  by  the  author  (559)  in  1913.  At  that  time, 
there  appeared  the  paper  of  Abderhalden  and  Lampe  (I.e.  25),  in 
which  they  formulated  the  "toxic"  theory  of  beriberi.  The  symp- 
toms of  the  disease  were  apparent  later  on  cooked  rice  than  on  raw 
rice.  This  observation,  which  in  itself  was  correct,  but  which  was 
thought  to  be  due  to  the  elimination  of  toxins,  may  be  explained  in  a 
very  simple  way.  Cooked  rice  takes  up  so  much  water  that  it  is 
impossible  to  feed  an  amount  corresponding  to  uncooked  rice,  and 
the  results  appear  as  follows : 


COOKED  RICE 

RAW  RICE 

Appearance  of  beriberi 

Dead 

Appearance  of  beriberi 

Dead 

days 

days 

days 

days 

44 

44 

25 

26 

However,  if  pigeons  were  fed  cooked  rice  corresponding  to  10  grams 
of  raw  rice,  then  the  results  were  somewhat  different : 


COOKED  RICE 


Appearance  of  beriberi 

Dead 

Appearance  of  beriberi 

Dead 

days 

27 

days 

29 

days 

28 

days 
30 

212 


THE    VITAMINES 


Here,  we  see  that  the  disease  develops  almost  simultaneously  when 
comparable  amounts  of  rice  are  fed.  Later,  we  (560)  showed  that 
when  pigeons  are  fed  5,  10  and  20  grams  of  rice  daily  that  the  disease 
develops  the  sooner,  the  more  rice  is  fed : 


AMOUNT  OF  RICE 

5  grams 

10  grams 

20  grams 

Beriberi  

days 

39 

days 

36 

days 
22 

Dead  .. 

42 

38 

22 

We  also  tested  this  observation  on  a  synthetic  diet,  the  pigeons 
receiving  equal  quantities  of  food  (12.5  grams)  in  every  case: 


DIET 

SALTS 

CASEIN 

SUGAR 

LARD 

STARCH 

BERIBERI 

A 

4 

60 

12 

12 

12 

30  days 

B 

4 

12 

12 

60 

12 

40  davs 

C 

4 

12 

12 

12 

60 

24  days 

D 

4 

12 

60 

12 

12 

'   28  days 

From  these  findings  we  concluded,  at  that  time,  .that  vitamine  B 
plays  a  particular  part  in  the  carbohydrate  metabolism,  since  on 
feeding  large  amounts  of  carbohydrates,  the  pigeons  developed  the 
disease  sooner.  Our  observations  on  the  influence  of  rice  feeding 
were  confirmed  by  Braddon  and  Cooper  (I.e.  551  and  561)  in  1914, 
by  Weill  and  Mouriquand  (562)  and  also  by  Green  (I.e.  526) .  Green, 
however,  assumed  that  not  only  carbohydrates  but  the  entire  metabo- 
lism plays  a  role  in  the  vitamine  requirements.  However,  other 
investigators  were  not  in  accord  with  our  views.  For  example, 
Eijkman  and  Hoogenhuyze  (I.e.  557)  found  that  an  addition  of 
meat  did  not  prevent  the  appearance  of  beriberi  symptoms.  Inas- 
much as  this  disease  may  occur  likewise  on  an  exclusive  meat  diet, 
they  used  this  in  combating  our  views.  Since  about  40  per  cent  of 
the  protein  in  the  body  is  transformed  into  glycogen,  and  since  the 
destruction  of  the  tissues  proceeds  similarly  in  starvation,  these 
facts,  in  reality,  cannot  be  used  to  discredit  our  findings.  Vedder's 
(563)  conclusions  were  similar  to  those  of  the  Dutch  investigators. 
Altogether  he  carried  out  five  experiments,  which  are  perhaps  less 
convincing  than  appears  at  first  sight. 


THE   ANTIBERIBERI   VITAMINE  213 

In  the  first  experiment  he  fed  11  chickens,  varying  in  weight  between  1800 
and  3800  grams  on  rice,  which  they  ate  voluntarily;  the  uneaten  rice  was 
weighed  the  next  day.  No  relationship  was  evident  between  the  appearance 
of  beriberi  and  the  amount  of  rice  utilized.  Since  the  birds  varied  consid- 
erably in  age  and  vitality,  so  that  the  energy  expended  and  required  were 
different,  and  since  this  experiment  was  carried  out  on  two  series  of  experi- 
mental animals,  in  tw.o  different  months,  with  a  possibility  of  the  dissimi- 
larity of  the  external  temperature,  therefore  this  experiment  is  not  quite 
significant.  In  a  second  experiment,  Vedder  fed  forcibly  100,  50,  and  25 
grams  of  white  rice  daily  to  chickens  averaging  1200  grams  in  weight.  Vedder 
then  stated  that  the  pigeons  could  die  as  well  from  over-feeding  as  from  under- 
feeding, because  rice,  collecting  in  the  crop,  mechanically  caused  disturbances 
not  related  to  the  stimulation  of  metabolism.  If  we  assume,  with  Vedder, 
that  chickens  fed  100  gm.  of  rice  really  die  as  a  result  of  over-feeding,  and 
only  consider  the  birds  receiving  less  rice,  then  chickens  on  50  grams  of  rice 
developed  beriberi  in  20  days  and  those  on  25  grams  of  rice,  much  later,  which 
naturally  completely  corroborates  the  results  of  other  investigators.  In  a 
third  experiment,  unpolished  rice  was  used,  but  in  this  case,  the  animals  did 
not  die  as  a  result  of  over-feeding,  although  they  were  given  50  and  75  grams 
of  rice  daily.  In  a  fourth  experiment,  chickens  were  fed  on  sterilized  meat 
and  eggs,  and  the  results  show  definitely  that  eggs  contain  some  vitamine 
while  meat  contains  a  greater  quantity.  Vedder  believed  that  if  beriberi 
could  occur  in  the  absence  of  carbohydrates,  then  it  was  certain  that  vitamine 
B  played  no  part  in  the  metabolism  of  this  dietary  component.  We  have 
already  seen  that  this  conclusion  is,  in  all  probability,  incorrect.  If  these 
foods  were  vitamine-free,  the  retardation  of  the  beriberi  symptoms  would  be 
the  best  demonstration  for  the  correctness  of  our  observations.  In  the  fifth 
and  last  experiment,  chickens  were  fed  on  sterilized  eggs  and  meat,  with  the 
addition  of  white  rice.  The  eggs,  apparently  egg-white  and  egg-yolk,  were 
evidently  not  entirely  vitamine-free,  and  the  residual  vitamine  might  perhaps 
suffice  for  25  grams  of  rice.  The  eggs  and  meat  used  here,  according  to  Vedder's 
statements,  were  supposed  to  be  identical  with  the  preparations  used  in  the 
fourth  experiment.  This  is  very  unlikely,  since  the  chickens,  in  the  fourth 
experiment,  developed  beriberi  in  61  days,  when  fed  on  50  grams  of  eggs, 
while  in  the  fifth  experiment,  the  animals  developed  no  beriberi  symptoms 
on  an  addition  of  25  grams  of  eggs.  As  we  see  from  the  above,  Vedder's  objec- 
tions do  not  carry  any  weight.  Aside  from  this,  upon  examining  our  pigeon 
experiments  more  closely  and  disregarding  those  fed  on  larger  quantities  of 
rice,  the  results  on  lesser  quantities  of  rice  still  prove  our  contention.  The 
amount  of  food  was  kept  constant  (12.5  grams  daily)  in  another  experiment, 
where  a  synthetic  diet  was  used  which  confirmed  our  rice  results,  so  that  the 
food  only  varied  qualitatively  but  not  quantitatively. 

The  chief  difficulty  encountered  in  this  experiment  was  that  the 
amount  of  vitamine  needed  for  metabolism  depends  not  on  the 
ingested  quantity  of  food,  but  on  the  portion  assimilated.  This  con- 


214  THE    VITAMINES 

dition  is  controlled  with  difficulty  in  pigeons  and  chickens,  because 
of  the  scattering  of  the  food  and  the  ease  with  which  they  vomit. 

It  would,  therefore,  be  desirable  to  repeat  these  experiments  on 
rats,  in  which  it  is  easy  to  control  the  food.  Such  experiments, 
which  were  carried  out  by  Funk  and  Dubin  (I.e.  331),  and  which  con- 
firmed in  general  the  results  with  pigeons,  will  be. described  in  another 
chapter.  Whether  our  view,  that  vitamine  B  plays  an  important 
role  in  carbohydrate  metabolism,  may  be  regarded  as  correct  must 
remain  for  the  future  to  show,  since  simultaneously  with  an  enrich- 
ment of  the  carbohydrates  in  the  form  of  starch  or  sugar,  there  is  a 
decrease  in  protein.  That  protein  plays  a  particular  role  in  nutrition, 
is  well  known.  Maignon  (564)  fed  rats  on  diets  which  varied  con- 
siderably in  their  protein,  starch  and  fat  content,  but  unfortunately 
the  vitamines  were  not  provided  for.  While  each  of  the  protein-fat 
combinations  could  maintain  the  animal  at  a  constant  weight  for  50 
days,  this  was  true  of  the  protein-starch  mixture  only  when  both 
components  were  present  in  equal  quantities.  From  this,  it  was  con- 
cluded that  protein  is  better  utilized  in  the  presence  of  fat  than  in 
that  of  starch.  The  best  nutrition  was  observed  when  a  mixture 
consisting  one  one  part  protein,  one  part  fat  and  1.33  parts  starch 
was  fed.  Bierry  and  Portier  (565)  obtained  similar  results,  and 
expressed  the  view  that  there  must  be  a  very  definite  relationship 
between  the  protein  and  fat  of  any  diet.  Bierry  (566)  noted  that 
disturbances  in  metabolism  could  be  prevented  only  when  the  individ- 
ual dietary  constituents  were  present  in  proper  proportions. 

While  we  believed,  at  first,  that  the  delaying  effect  of  a  protein 
addition  on  the  development  of  beriberi  was  due  to  the  presence 
therein  of  vitamine  B,  this  view  is  perhaps  no  longer  tenable.  We 
shall  see  later  that  the  "vitamine  sparing"  action  of  protein,  or  the 
presence  of  a  special  substance  essential  to  life  may  play  a  part  in 
this  phenomenon.  For  this  reason,  we  must  be  conservative  with 
our  conclusions  when  extra  protein  is  added  to  the  diet.  To  illustrate, 
let  us  consider  the  work  of  Johns  and  Finks  (567),  who  investigated 
the  nutritive  value  and  B-vitamine  content  of  war  bread,  which 
was  milled  to  the  extent  of  7'4  per  cent.  Working  with  rats,  they 
found  that  the  above  food  contains  sufficient  B-vitamine  but  not 
enough  protein,  since  the  animals  resumed  normal  growth  on  the 
addition  of  casein.  This  conclusion  is  perhaps  not  quite  correct, 
for  the  growth-inhibiting  influence  may  possibly  still  have  been  due 


THE   ANTIBEKIBERI   VITAMINE  215 

to  a  lack  of  vitamine  B.  Similar  conditions  are  frequently  met 
with  in  the  study  of  pellagra  and  hunger  edema,  to  which  we  shall' 
refer  again  in  the  chapter  dealing  with  the  question.  We  have 
already  seen  in  the  investigation  of  Funk  and  Dubin  (I.e.  331)  that 
a  larger  addition  of  carbohydrates  increased  the  vitamine  B  require- 
ments. In  this  respect,  rats  are  similar  to  pigeons  except  that  in 
rats,  an  excess  of  fat  causes  much  greater  disturbances  than  in 
pigeons. 

Besides  the  above  mentioned  results,  Funk  and  v.  Schonborn  (I.e. 
304)  found  that  whereas  in  normal  pigeons,  the  glycogen  and  blood 
sugar  content  of  the  liver  is  1.17  per  cent  and  0.1  per  cent,  respec- 
tively, on  an  artificial  vitamine-free  diet,  the  glycogen  content  drops 
to  0.48  per  cent  and  the  blood  sugar  rises  to  0.15  per  cent.9  Pigeons 
fed  on  an  excess  of  sugar  showed  4.5  per  cent  glycogen  and  0.15  per 
cent  blood  sugar.  On  an  excess  of  starch,  there  is  no  glycogen  and 
0.26  per  cent  blood  sugar.  In  the  last  case,  if  vitamine  B  is  added, 
the  values  for  glycogen  and  blood  sugar  approach  normal.  These 
results  were  later  repeated  several  times  and  corroborated  by  the 
author  (568). 10 

Besides  this,  we  also  studied  the  influence  of  substances  having  a 
known  action  on  carbohydrate  metabolism.  Among  them,  the 
effect  of  glucose,  phlorizin,  adrenaline,  pituitrin,  thyroid  and  para- 
thyroids on  pigeon  beriberi  was  tested,  normal  and  rice-fed  pigeons 
being  used.  Of  special  interest,  was  adrenaline,  which  shortened  the 
life  of  the  animal;  this  was  also  true  of  thyroid.  The  influence  of 
parathyroids  was  quite  different.  While  thyroid  raised  the  blood 
sugar  content  and  the  arnino  nitrogen  in  the  serum,  and  lowered  the 
glycogen  content  of  the  liver,  the  addition  of  parathyroids  increased 
the  glycogen  and  amino  acid  content,  but  had  little  effect  on  blood 
sugar.  We  must  admit,  however,  that  Elias  and  Kolb  (569)  also 
demonstrated  glucosuria  in  starving  dogs,  which  was  attributed 
to  acidosis,  and  which  could  be  remedied  by  the  administration  of 
alkali.  We  have  already  emphasized  that  it  is  not  known,  at  present 
how  many  of  the  starvation  symptoms  may  be  regarded  as  results  of 

*  Scott  (567a)  has  found  the  normal  blood  sugar  content  in  pigeons  higher, 
but  the  difference  may  be  due  to  a  different  method  of  bleeding  the  animal, 
a  fact  which  has  been  frequently  observed  by  us. 

10  The  finding  of  glucosuria  in  pigeon  beriberi  has  not  yet  been  confirmed 
in  human  beriberi. 


FIG.  51.  COMPARISON  OF  THE  ADRENALINE  CONTENT — TESTED  ON  SHEEP — OP 

THE  SUPRARENALS  OF  PlGEONS  IN  BERIBERI,   NORMAL 

NUTRITION  AND  INANITION 

I,  normal  pigeons;  II,  injection  of  0.01  mgm.  adrenaline  into   sheep   as 
control.  Ill,  inanition;  IV  beriberi  (McCarrison) . 

216 


THE    ANTIBEKIBERI   VITAMINE  217 

an  avitaminosis,  and  how  many  of  the  beriberi  symptoms  must  be 
attributed  to  a  condition  of  starvation. 

The  discovery  of  glucosuria  in  pigeon  beriberi  may  evidently  be 
connected  with  some  observations  made  by  McCarrison  (I.e.  296). 
He  found  that  in  avian  beriberi,  the  suprarenals  undergo  considerable 
enlargement  and  are  characterized  by  a  high  adrenaline  content. 
Here  too,  an  increased  adrenaline  content  was  found  in  starving 
pigeons,  but  not  so  much  as  in  beriberi  pigeons.  The  adrenals  were 
also  somewhat  enlarged.  In  the  same  publication  we  find  numerous 
data  on  temperature,  respiration  and  weight  of  the  individual  organs 
in  avian  beriberi,  which  are  also  of  physiological  interest. 

In  concluding  this  chapter,  we  wish  to  point  out  that  in  considering 
the  vitamine  B  requirements,  we  must  take  note  of  the  composition 
of  the  diet  as  regards  protein,  carbohydrate  and  fat.  These  require- 
ments are  possibly  dependent  also  on  the  simultaneous  presence  or 
absence  of  other  vitamines.  If,  for  example,  the  quantity  of  vitamine 
A  is  too  little,  the  requirements  for  vitamine  B  are  perhaps  increased. 
However,  there  are  as  yet  no  available  data  on  this  point. 


CHAPTER  II 
THE  ANTIRACHITIC  VITAMINE — VITAMIN E  A 

The  history  of  the  discovery  of  vitamine  A  has  already  been  dis- 
cussed, so  that  we  may  take  up  the  chemistry  of  this  substance  at 
once.  We  have  noted  that  for  a  long  time,  vitamine  A  was  supposed 
to  be  free  from  nitrogen,  but  we  feel  that  in  the  interest  of  future 
investigation  this  question  should  be  left  open.  We  must  also  leave 
for  the  future,  the  question  as  to  whether  the  designation,  vitamine 
A,  includes  several  substances  of  the  same  type,  or  only  a  single 
substance.  The  chemistry  of  vitamine  A  has  advanced  but  little 
so  that  we  shall  only  discuss  its  resistance  to  physical  agents,  solu- 
bility and  occurrence. 

CHEMISTRY    OF   COD   LIVER    OIL 

Cod  liver  oil  is  the  only  starting  material  from  which  attempts 
have  been  made  to  isolate  the  vitamine.  Although  Gsborne  and 
Mendel  (570)  showed  in  1914  that  this  oil  contains  vitamine  A,  and 
although,  because  of  its  relationship  to  the  therapy  of  rickets,  it  has 
commanded  attention  for  a  long  time,  we  are  still  in  the  dark  as  to 
the  nature  of  the  vitamine  contained  therein.  However,  when  we 
consider  the  known  facts  about  the  resistance  of  vitamine  A  from 
various  sources,  and  the  technical  preparation  of  cod  liver  oil  (571), 
it  is  obvious  that  because  of  the  stability  of  the  vitamine,  cod  liver 
oil  is  the  best  material  to  use  for  the  chemical  isolation  of  vitamine  A. 
Iscovesco  (572)  believed  that  almost  all  of  the  phosphorus  and  nitro- 
gen of  cod  liver  oil  occurs  as  a  lecithide  or  as  a  lecithalbumen.  If 
acetone  is  added  to  cod  liver  oil,  with  stirring,  a  precipitate  is  formed , 
which,  on  reaching  a  maximum,  is  redissolved.  The  oil,  freed  from 
some  of  its  lecithin  by  acetone,  was  shaken  with  an  equal  volume  of 
95  per  cent  alcohol.  To  the  alcoholic  solution,  which  contains  all 
of  the  phosphorus  of  the  oil,  absolute  alcohol  was  added,  giving  two 
separate  fractions.  The  soluble  part  was  concentrated  at  a  low 
temperature,  the  residue  dissolved  in  ether  and  precipitated  with 
acetone.  In  this  way,  from  1  kilo  of  oil,  0.02  gram  of  the  pure 
lecithide  was  obtained,  which  was  soluble  in  benzol  and  chloroform, 

218 


THE   ANTIKACHITIC   VITAMINE  219 

melted  at  70°C.,  and  contained  60  per  cent  C,  7.3  per  cent  H,  2.1 
per  cent  N  and  4  per  cent  P.  A  part  of  this  semi-solid  mass  was 
used  in  an  experiment  on  rabbits  kept  on  a  normal  diet.  The  leci- 
thide  in  olive  oil  was  injected  subcutaneously  every  second  day  for 
130  days.  The  result  were  as  follows : 

Gain  in  weight 
per  cent 

Controls 33 

Cod  liver  oil 55 

Cod  liver  oil  without  lecithide 37 

Olive  oil 33 

Olive  oil  with  0.5  per  cent  lecithide 56 

It  would  appear  from  these  results  that  the  total  active  substance 
was  present  in  the  lecithide,  but  it  is  still  necessary  to  corroborate 
these  findings  in  rats  fed  on  a  diet  free  from  vitamine  A. 

Other  investigators  turned  their  attention  to  the  nitrogenous  por- 
tion of  the  oil.  In  1888,  Gautier  and  Mourgues  (573)  showed  that 
even  a  purified  oil  contains  a  small  amount  of  organic  bases  which 
at  that  time  were  thought  to  be  ptomaines  or  alkaloids. 

One  hundred  kilos  of  oil  were  treated  with  an  equal  volume  of  33  per  cent 
alcohol,  to  which  4  grams  oxalic  acid  per  liter  were  added.  The  watery- 
alcoholic  solution  was  saturated  with  calcium  hydroxide,  filtered  and  the 
filtrate  evaporated  in  vacuum  at  45°C.  At  the  end  of  the  distillation,  calcium 
carbonate  and  hydrate  were  added,  the  mixture  evaporated  to  dryness  aud 
the  residue  extracted  with  90  per  cent  alcohol.  To  this  extract,  after  the 
removal  of  the  alcohol  in  vacuum,  concentrated  potassium  hydroxide  was 
added  and  the  alkaline  solution  extracted  with  ether.  An  ether  solution  of 
oxalic  acid  was  then  added,  to  precipitate  the  bases  present  as  oxalates.  The 
yield  was  52  to  65  grams  oxalate  from  100  kilos  of  oil.  The  oxalates  were  dis- 
solved in  dilute  potassium  hydroxide,  and  the  bases  set  free  in  the  form  of  an 
oil,  which  was  dried  and  fractionated,  giving  the  following  fractions: 

Fraction  I,  87-90°C.  butylamine;  fraction  II,  96-98°C.  amylamine;  fraction 
III,  under  100°C.  hexylamine;  fraction  IV,  between  198-200°C.  hydrotoluidine. 
The  distillation  was  then  continued  to  215°C.  whereupon  the  brownish  distil- 
lation residue  was  extracted  with  ether.  The  ether  solution  was  evaporated, 
the  residue  treated  with  HC1  and  precipitated  with  a  solution  of  platinic 
chloride,  giving  a  base  as  a  double  salt,  called  "aselin,"  having  the  compo- 
sition C25H32N4.  From  the  mother  liquor,  a  second  base  was  isolated,  called 
"morrhuin,"  having  the  formula  dgH^Ns.  It  is  not  impossible  that  both  of 
these  bases  were  rather  condensation  or  decomposition  products,  arising 
secondarily  during  the  distillation. 

This  work,  which  is  of  little  more  than  historical  interest,  shows 
nevertheless  that  cod  liver  oil  contains  nitrogenous  substances  which 
are  of  interest  to  us  because  of  the  vitamine  A. 


220  THE   VITAMINES 

The  author  (574)  has  investigated  a  crude  oil  and  found  that  the 
quantity  of  extractives  therein  was  much  greater  than  in  the  purified 
oil.  [Crude  oil  was  found  more  active  than  refined  oil  in  animal 
experiments  by  Zilva  and  Miura  (574a)].  23.5  kilos  of  the  oil  were 
extracted  three  times  with  about  660  cc.  alcohol,  2  liters  of  water  and 
50  cc.  concentrated  HC1;  the  extracts  were  concentrated  in  vacuum, 
the  oily  residue  hydrolyzed  with  sulphuric  acid  and  precipitated 
with  phosphotungstic  acid.  The  practically  dry  precipitate  weighed 
969  grams.  The  oil,  hydrolyzed  with  sulphuric  acid  (2  hours)  gave 
an  additional  186  grams  of  this  precipitate,  while  the  oil  remaining 
after  the  concentration  of  the  water-alcohol  solution  gave  only  19.5 
grams  of  the  precipitate.  A  second  portion  of  oil,  weighing  25 
kilos  and  treated  with  sulphuric  instead  of  hydrochloric  acid,  gave 
1117  grams  phosphotungstate,  of  which  61.2  grams  was  insoluble  in 
acetone.  This  fraction,  after  the  decomposition  with  lead  acetate, 
gave  a  precipitate  with  sublimate  in  alcoholic  solution.  A  great 
number  of  fractions  were  prepared  by  us,  but  could  not  be  tested  for 
various  reasons. 

One  of  the  common  conceptions  regarding  cod  liver  oil  was  that 
the  mysterious  action  of  unsaturated  oils  was  ascribed  to  it.  How- 
ever, this  view  has  not  yet  been  experimentally  tested.  Paal  and 
Roth  (575)  reduced  cod  liver  oil  with  colloidal  palladium  and  hydro- 
gen to  a  solid,  whereby  the  reduction  of  all  the  unsaturated  bonds 
was  almost  completely  quantitative.  The  general  nutritive  value 
of  such  an  oil  was  investigated  by  Suzuki  and  his  co-workers  (I.e.  323) 
and  found  favorable.  Chapman  (576)  and  a  number  of  Japanese 
workers  including  Tsujimoto  (577)  and  Kubota  (578)  isolated  from 
cod  liver  oil  a  very  unsaturated  hydrocarbon  called,  "spinacen"  or 
"squalen,"  which,  however,  was  found  valueless  as  a  substitute  for 
vitamine  A  by  Suzuki.  Bull  (579)  obtained  a  mixture  of  C2oH3oBrio02 
and  C2oH28Bri202  on  brominating  cod  liver  oil.  For  use  in  the 
therapy  of  leprosy,  such  products  as  sodium  morrhuate  were  prepared 
by  Rogers  (580). 

CHEMISTRY   OF   VITAMINE   A   IN   BUTTER   AND    OTHER   SOURCES 

The  data  on  the  chemistry  of  this  vitamine  in  butter  are  very 
uncertain  and  not  infrequently  contradictory.  The  first  experiments 
on  the  isolation  of  vitamine  A  from  butter  were  made  by  McCollum 
and  Davis  (I.e.  82).  They  reported  that  when  a  solution  of  butter 


THE   ANTIKACHITIC   VITAMINE  221 

\ 

in  petroleum  ether  is  hydrolyzed  with  alcoholic  potassium  hydroxide 
at  ordinary  temperature,  and  the  soap  solution  neutralized,  the  active 
substance  may  be  concentrated  in  olive  oil  by  extracting  with  the 
latter.  Osborne  and  Mendel  (581)  dissolved  ox  fat  and  butter  in 
alcohol  at  40°C.  and  allowed  it  to  crystallize  out  on  freezing  at 
—  20°C.  The  A-vitamine  could  not  be  demonstrated  in  the  crystal- 
lized portion,  but  was  found  in  the  oily,  uncrystallizable  portion. 
Drummond  (582)  attempted  to  repeat  the  work  of  McCollum  and 
Davis  on  the  stability  after  saponification.  After  butter  had  been 
saponified  at  the  usual  temperature,  the  solvent  was  removed  in 
vacuum,  the  soaps  dissolved  in  water  and  the  unsaponified  portion 
extracted  with  ether.  All  of  the  fractions  thus  obtained  were  tested 
on  rats  and  found  inactive.  Recently,  Steenbock,  Sell  and  Buell 
(582a)  repeated  the  saponification  experiments  and  have  found 
that  cod  liver  oil  can  be  treated  with  20  per  cent  alcoholic  potassium 
hydroxide  solution  for  4  hours  at  37°C.  without  losing  its  potency. 
Since  the  active  substance  after  dilution  with  water  can  be  extracted 
with  ether,  it  was  concluded  that  vitamine  A  is  neither  a  fat  nor 
an  ester. 

Numerous  experiments  were  carried  out  for  the  purpose  of  con- 
centrating this  vitamine  from  various  vegetables  and  grasses  by 
extraction.  McCollum,  Simmonds  and  Pitz  (583)  thought  that  the 
vitamine  could  not  be  removed  by  extracting  plant  tissue  with  ether, 
because  it  is  in  all  probability  bound  to  the  protein.  Still,  Osborne 
and  Mendel  (584)  were  able  to  do  this.  They  dried  spinach  and 
clover  in  a  stream  of  air  at  60°C.,  and  then  extracted  it  with  ether, 
obtaining  a  very  active  substance.  These  results  were  confirmed  by 
Zilva  (585)  who  obtained  an  active  alcoholic  and  ether  extract  from 
carrots.  Steenbock  and  Boutwell  (586)  were  unable  to  obtain  an 
active  substance  by  extraction  of  carrots  with  ether  alone,  but  could 
do  so  with  carbon  disulphide,  chloroform  and  benzol.  In  spite  of 
this,  the  residues  showed  almost  the  original  activity.  Using  alfalfa 
grass  as  a  source,  a  simple  ether  extraction  gave  no  result,  while  a 
mixture  of  alcohol  and  benzol  gave  almost  quantitative  extraction. 
The  same  workers  then  tried  to  make  a  fractionation  by  means  of 
solubility.  Alfalfa  was  treated  with  cold  alcohol,  the  latter  distilled 
off  on  the  water  bath  and  the  residue  allowed  to  stand  over  night 
with  20  per  cent  alcoholic  potassium  hydroxide.  When  the  saponifi- 
cation had  been  treated  with  ether  and  filtered  from  the  ether- 


222  THE   VITAMINES 

insoluble  portion,  the  ether  solution,  after  washing  with  water,  was 
shown  to  be  active.  From  this  residue,  the  carrotin  was  separated 
from  xanthophyll  by  extraction  with  petroleum  ether,  showing  the 
carrotin  fraction  to  be  active  and  the  xanthophyll  fraction,  inactive. 

THE   NATURE    OF   VITAMINE   A 

Drummond  (I.e.  582)  in  one  of  his  papers  proposed  the  conception 
that  vitamine  A  was  of  the  nature  of  a  ferment  and  was  a  more  or 
less  unobtainable  body.  He  carried  out  experiments  with  various 
known  substances  and  found  that  fatty  acids,  glycerol,  cholesterol, 
lecithin,  sphingosine,  phrenosine,  kephalin,  lipochrome  and  carrotin 
could  not  simulate  the  action  of  vitamine  A.  Steenbock  (587)  made 
the  interesting  observation  that  the  vitamine  A  content  of  maize 
goes  parallel  with  the  content  of  yellow  pigment;  white  maize  was 
found  free  from  this  vitamine,  and  the  same  was  also  true  of  most 
kinds  of  vegetables.  According  to  Steenbock  and  Boutwell  (588) 
when  white  corn  is  supplemented  by  butter,  clover  or  spinach,  the 
rat  resumes  growth.  Palmer  (589),  on  the  contrary,  was  opposed 
to  this  finding.  In  the  first  place,  he  showed  that  chickens  could 
live  on  a  diet  free  from  carrotinoids,  at  least  through  one  generation, 
although  it  is  of  course  quite  possible  that  chickens  may  live  without 
vitamine  A,  or  at  best  with  very  little.  Second,  it  was  possible  for 
him  to  raise  mammals  (rats;  Palmer  and  Kennedy,  589a),  the  blood 
and  subcutaneous  tissue  of  which  were  pigment  free.  Despite  the 
fact  that  the  milk  of  these  animals  was  practically  pigment-free, 
it  was  possible  for  them  to  suckle  and  rear  their  young.  Palmer 
observed  also  that  there  are  oils,  such  as  cottonseed  oil,  which, 
although  containing  pigment,  show  a  lack  of  vitamine  A.  Rosen- 
heim  and  Drummond  (590)  took  up  this  question  and  in  general 
were  able  to  confirm  the  fact  that,  as  a  rule,  foodstuffs  containing 
lipochrome  also  contain  vitamine  A.  Still,  it  was  found  that  a 
pure  carrotin  solution  is  free  from  this  vitamine,  and  the  authors 
believed  that  although  the  latter  must  be  different  from  carrotin 
and  xanthophyll,  it  is  nevertheless  associated  with  these  pigments. 
As  to  what  action  chemically  pure  carrotin  has  on  rats,  opinions 
differ.  While  Stephenson  (590a)  finds  that  pure  carrotin  (m.p. 
172-173°C.)  is  entirely  inactive  and  while  butter  decolorized  with 
charcoal  and  free  from  pigment  is  still  active,  Steenbock,  Sell, 
Nelson  and  Buell  (590b)  have  obtained  normal  growth  with  pure 


THE   ANTIRACHITIC   VITAMINE  223 

carrotin.  They  prepared  vitamine  A  from  alfalfa  and  found  that 
it  withstood  saponification  with  heat,  oxidation  with  hydrogen 
peroxide  and  reduction  with  nascent  hydrogen.  From  the  unsaponi- 
fiable  fraction,  active  crystalline  acetylated  products  have  been 
prepared.  Liver  tissue  contains,  besides  the  two  known  yellow 
pigments,  another  substance  which  gives  the  lipochrome  reaction 
and,  in  addition,  the  color  reaction  with  sulphuric  acid  which  is 
regarded  in  the  literature  as  being  specific  for  cod  liver  oil.  This 
substance  is  not  a  casual  impurity  as  it  may  always  be  demon- 
strated in  cod  liver  oil  and  in  liver  fats.  Hi j  mans  van  den  Bergh 
and  Muller  (591)  have  described  methods  which  may  be  used  for 
the  separation  of  the  lipochromes  in  blood.  In  order  to  liberate 
them,  it  was  necessary  to  precipitate  the  serum  proteins  with  alcohol. 
It  was  assumed  that  other  chemical  substances  were  associated  with 
the  pigments.  Drummond  and  Coward  (592)  have  investigated 
the  pigment  and  vitamine  A  content  of  a  number  of  oils  and  fats, 
and  found  that  these  two  factors  do  not  always  occur  together.  In 
butter,  the  pigment  content  was  found  to  run  somewhat  parallel  to 
that  of  vitamine  A,  whereas  certain  pork  fats  and  dog  fat  contain 
no  pigment,  but  a  marked  amount  of  vitamine.  They  came  to  the 
conclusion  that  a  relationship  between  the  pigment  and  vitamine  A 
could  not  be  maintained  unless  we  assume,  with  Steenbock,  that  in 
some  cases  the  pigments  occur  in  the  leuco-form.  As  evidenced  by 
the  above,  the  whole  question  still  rests  on  an  uncertain  foundation 
and  can  be  solved  only  when  a  purification  of  this  substance  is  made. 
Properties  of  vitamine  A .  The  characteristics  of  this  vitamine  are 
as  yet  very  little  known.  As  for  the  question  whether  the  substance 
is  soluble  in  water,  it  must  remain  open  for  the  present.  McCollum, 
Simmonds  and  Steenbock  (593)  have  stated  that  fat-free  milk  still 
contains  appreciable  amounts  of  vitamine  A,  a  finding  which  seems 
to  be  in  accord  with  the  latest  results.  According  to  their  work,  the 
vitamine  may  be  extracted  with  small  quantities  of  water  from  melted 
butter  by  repeated  extraction,  the  remaining  butter  being  inactive. 
On  the  other  hand,  Steenbock,  Boutwell  and  Kent  (594)  showed  that 
both  the  residual  butter  and  the  watery  extract  were  inactive.  They 
believed  that  heat  and  not  oxidation  is  the  reason  for  the  destruction, 
although  we  shall  see  that  there  is  still  another  complication  to  be 
considered.  The  finding  of  McCollum,  Simmonds  and  Steenbock 
(I.e.  593)  that  fat-free  skimmed  milk  still  contains  vitamine  A  was 
again  confirmed  by  Hopkins  (595). 


224  THE    VITAMINES 

The  first  statement  on  the  stability  of  the  active  substance  from 
butter  was  made  by  Osborne  and  Mendel  (I.e.  581).  They  found 
that  when  steam  was  passed  through  butter  for  2J  hours,  the 
vitamine  was  unaffected.  That  this  substance  in  egg-yolk  is  resistant 
to  heat,  was  shown  by  McCollum  and  Davis  (596).  In  this  connec- 
tion, Drummond  (597)  found  that  vitamine  A  from  butter  and  other 
sources  is  very  unstable  to  heat,  thus  corroborating  the  statements  of 
Steenbock.  Drummond  also  reported  that  oxidation  does  not  seem 
to  play  any  part  in  the  destruction  of  this  substance.  Steenbock  and 
Boutwell  (598)  found  yellow  corn  and  alfalfa  active  after  heating  in 
the  autoclave  for  three  hours.  Osborne  and  Mendel  (599)  found 
butter,  as  such,  much  more  stable  against  storage  and  the  influence  of 
light,  than  the  uncrystallizable  part — butter  oil. 

All  of  these  apparently  conflicting  statements  may  be  well  under- 
stood when  the  new  observation  of  Hopkins  (I.e.  595  and  599a) 
is  taken  note  of.  He  found  that  in  the  destruction  of  vitamine  A 
oxidation,  not  temperature,  plays  the  chief  part.  This  has  been 
confirmed  by  Drummond  and  Coward  (599b)  and  is  in  agreement 
with  the  observation  of  Osborne  and  Mendel  (600),  who  found  butter 
stable  when  steam  was  passed  through  it,  but  not  when  the  butter 
was  heated  in  the  absence  of  water.  The  stability  in  this  case  may 
be  explained  by  the  replacement  of  air  by  steam,  although  even  dry 
butter  could  be  heated  at  96°C.  for  15  hours  without  noting  any  loss 
of  activity. 

As  regards  the  action  of  physical  factors,  Zilva  (I.e.  513)  showed 
that  butter  lost  its  activity  on  exposure  to  ultra-violet  rays  for  8 
hours.  The  possibility  of  the  formation  of  ozone  was  considered  in 
a  later  paper,  and  it  was  actually  found  that  ozone  is  destructive 
to  vitamine  A  (600a). 

Although  the  destruction  of  vitamine  A  by  oxidation  has  been  well 
explained  by  various  statements,  it  is  more  difficult  to  understand 
the  destruction  of  fat  on  reducing  at  a  temperature  of  75°  to  120°C., 
as  carried  out  by  Fahrion  (601).  For,  in  the  first  place,  vitamine  A, 
according  to  the  newer  findings,  is  quite  resistant  to  heat,  and 
second,  the  substances  sensitive  to  oxidation  are,  for  the  most  part, 
resistant  to  reduction.  Nevertheless,  according  to  the  findings  of 
Drummond  and  Coward  (I.e.  592),  as  well  as  of  Fahrion,  the  reduced 
fats  are  totally  inactive.  Fahrion  believed  that  vitamine  A  is  almost 
insoluble  in  fat  solvents. 


THE   ANTIRACHITIC   VITAMINE  225 

Considering  the  manner  of  preparing  commercial  cod  liver  oil, 
we  must  marvel,  in  the  light  of  the  above  mentioned  observations, 
that  the  oil  still  has  therapeutic  properties.  In  this  connection,  it 
would  be  interesting  to  observe  the  action  of  oxy-cod  liver  oil — a 
preparation  which,  according  to  Freudenberg  and  Klocman  (602), 
is  made  by  oxidation  of  cod  liver  oil  with  hydrogen  peroxide — as  well 
as  cod  liver  oil  emulsions  prepared  by  shaking  in  alkaline  medium. 
The  influence  of  such  a  procedure  on  the  therapeutic  action  is  dis- 
cussed by  Hess  and  linger  (603).  If  the  vitamine  A  of  cod  liver  oil 
really  withstands  the  above  mentioned  treatment  and  long  storage, 
then  we  must  assume  that  the  vitamine  from  this  source  is  exception- 
ally stable,  a  characteristic  which  should  be  significant  in  further 
studies.  According  to  Zilva  and  Miura  (I.e.  574a),  cod  liver  oil 
contains  250  times  as  much  vitamine  A  as  butter,  and  3  milligrams 
of  this  oil  per  day  is  sufficient  for  rats.  Drummond,  Coward  and 
Watson  (603a),  found  that  certain  samples  of  butter  contained 
no  more  vitamine  A  than  refined  vegetable  oils.  The  above  data 
undoubtedly  explain  the  differences  observed  in  the  value  between 
cod  liver  oil  and  butter  in  the  treatment  of  rickets. 

DEMONSTRATION   OF   VITAMINE   A 

Till  now,  the  only  method  used,  as  we  have  already  seen,  is  the 
behavior  of  the  rat.  To  make  the  demonstration  definite,  it  is  neces- 
sary, according  to  Drummond  and  Coward  (604)  to  provide,  above  all, 
that  the  basal  diet  is  completely  free  from  this  substance.  The  basal 
diet  used  for  this  purpose  is  composed  as  follows : 

Purified  casein 18      Yeast  extract 5 

Purified  rice  starch 52      Orange  juice 5 

Purified  reduced  vegetable  oil . .  15      Salt  mixture 5 

The  casein  is  heated  in  open  dishes  for  24  hours  at  105°C.  Rice 
starch  has  been  shown  to  be  pure  for  this  purpose  and  may  be  used, 
as  such.  Completely  reduced  vegetable  oil,  usually  cotton  seed 
oil,  or  steam  distilled  palm -kernel  oil  (Stammers,  604a)  is  used  as 
a  fat.  If  one  is  uncertain  as  to  the  freedom  of  this  oil  from  vitamine 
A,  then  the  addition  of  fat  should  be  omitted  entirely.  Orange 
juice  is  free  from  vitamine  A^  while  yeast  extract  has  not  yet  been 
investigated  sufficiently  in  this  respect.  Rats  weighing  50  to  70 
grams,  and  4  to  5  weeks  old  should  show  no  growth  at  all,  on  this 


226  THE    VITAMINES 

diet.  If  growth  ensues,  it  may  be  concluded  that  the  basal  diet 
was  insufficiently  purified.  Older  animals,  weighing  over  100  grams 
should  not  be  used  since  the  vitamine  A  requirement  diminishes 
with  age.  If  the  growth  remains  stationary  during  10  to  14  days, 
the  substance  to  be  tested  may  be  added,  preferably  apart  from  the 
diet.  If  growth  is  seen,  it  may  be  concluded  that  vitamine  A  was 
present  in  the  substance  added. 

Significance  of  vitamine  A  in  physiology  and  pathology.  From  a 
study  of  this  vitamine  in  various  nuts,  Coward  and  Drummond  (605) 
came  to  the  conclusion  that  it  occurs  only  in  small  amounts  in  seeds, 
but  that  during  the  process  of  germination  it  appears  in  greater 
quantities.  We  have  already  mentioned  the  possibility  that  green 
leaves  may  synthetize  vitamine  A  under  the  influence  of  sunlight, 
and  in  this  connection  Delf  (605a)  has  found  that  the  inner  white 
leaves  of  cabbage  are  free  from  this  vitamine,  while  the  outer  green 
leaves  contain  the  substance.  Recently,  Coward  and  Drummond 
(605b)  have  experimentally  substantiated  these  findings,  showing 
that  green  leaves  form  vitamine  A  only  in  the  presence  of  sunlight, 
while  the  lower  organisms  devoid  of  chlorophyll  are  unable  to  syn- 
thetize this  vitamine. 

Hughes  (605c)  fed  a  number  of  animals  on  diets  rich  and  poor  in 
vitamine  A,  but  on  examination  of  the  organs,  no  relation  could  be 
shown  between  the  vitamine  A  content  of  the  food  and  that  of  the 
adipose  tissue.  Drummond,  Golding,  Zilva  and  Coward  (605d) 
explain  the  poverty  of  pig's  fat  in  vitamine  A  in  two  ways  (1)  inade- 
quacy of  the  diet  in  this  substance,  and  (2)  losses  by  oxidation  during 
the  melting  out  process  of  the  fat. 

We  have  already  stated  that  the  lack  of  vitamine  A  in  rats  is  fol- 
lowed by  no  characteristic  pathological  changes,  and  hence  it  is 
difficult  to  point  out  just  what  is  the  importance  of  this  substance. 
Drummond  (606)  tried  to  solve  this  question.  Since  this  substance 
is  associated  with  fats,  as  a  rule,  it  was  easiest  to  assume  that  it 
played  a  part  in  the  fat  metabolism,  but  it  was  not  possible  to  find 
such  a  relationship.  Rats  lacking  both  this  vitamine  and  fat  showed 
no  different  behavior  from  those  animals  lacking  only  one  of  these 
constituents.  The  temperature  of  these  rats  was  normal,  so  that 
this  substance  has  nothing  to  do  with  the  regulation  of  body  tem- 
perature. In  another  experiment,  Drummond  tried  to  see  whether 
or  not  vitamine  A  plays  a  role  in  the  synthesis  of  neutral  fats  from 


THE   ANTIRACHITIC   VITAMINE  227 

fatty  acids.  For  this  purpose,  one  series  of  animals  were  fed  with 
fat,  another  with  fatty  acids.  When  vitamine  A  was  absent  from 
the  diet,  the  fatty  acid  mixture  was  well  assimilated,  and  the  animals 
showed  a  normal  fat  content.  In  our  experiments,  together  with 
Dubin  (I.e.  331),  we  fed  rats  on  a  diet  containing  a  large  amount  of 
neutral  fat.  These  rats,  in  spite  of  the  administration  of  vitamine 
B,  ceased  growing  and  lost  markedly  in  body  weight.  We  tried  to 
improve  the  condition  of  the  animals  by  adding  greater  amounts  of 
vitamine  A  in  the  form  of  cod  liver  oil  or  butter,  but  without  success. 
Unlike  vitamine  B,  vitamine  A,  given  parenterally,  is  without  any 
action. 

The  role  of  vitamine  A  in  pathology  was  treated  by  Mendel  (607). 
It  was  particularly  noted  that  this  substance  was  of  less  importance 
for  adults  than  for  young  growing  animals.  It  was  also  shown  to 
be  of  significance  in  the  manifestation  of  ophthalmia,  as  well  as 
in  the  occurrence  of  urinary  calculi,  conditions  which  arise  chiefly 
when  vitamine  A  is  absent  from  the  diet.  McCarrison  (608)  holds 
that  vitamine  A  plays  a  part  in  the  prophylaxis  of  edema.  All  of 
these  statements  will  be  discussed  in  relation  to  each  other  in  another 
chapter. 


CHAPTER  III 

THE  RELATIONSHIP  OF  THE  ANTIBERIBERI  AND  THE  ANTIRACHITIC 

VlTAMINES   TO   LlPOIDS 

The  object  of  this  chapter  is  to  show  that  vitamines  B  and  C  have 
presumably  not  the  slightest  relation  to  the  lipoids.  It  is,  of  course, 
more  difficult  to  demonstrate  this  as  regards  vitamine  A.  The  asso- 
ciation of  these  two  classes  of  substances,  the  lipoids  and  the  vitam- 
ines, came  about  through  the  conception  that  lipoids,  and  especially 
lecithin,  are  not  pure  products  but  may  contain  vitamine  as  an 
impurity.  It  has  often  been  stated  by  Cronheim  (609),  and  by 
others,  that  lecithin  is  of  significance  in  metabolism,  while  Robertson 
(610)  noted  only  an  inhibition  of  growth  on  feeding  this  substance 
to  mice. 

In  biochemistry,  the  idea  has  long  been  prevalent  that  when 
animal  or  plant  tissue  is  extracted  with  alcohol,  or  better  still,  with 
ether,  the  fraction  in  the  solvent  can  be  regarded  as  a  lipoid.  In  our 
first  investigation,  carried  out  in  the  Lister  Institute  in  1911,  which 
had  for  its  purpose  the  isolation  of  vitamine,  we  convinced  ourselves 
that  an  extraction  with  alcohol  removes  a  great  deal  of  nitrogenous 
matter  (purines,  pyrimidines,  etc.),  in  which  the  vitamines  are  also 
to  be  found.  This  observation  was  made  use  of  by  McLean  (611), 
working  in  the  same  Institute  at  that  time,  to  remove  nitrogenous 
impurities  from  lecithin.  For  this  purpose,  a  crude  lipoid  from  horse 
meat  was  finely  divided  and  repeatedly  extracted  with  water.  In  the 
first  alcoholic  extract,  as  well  as  in  the  watery  extracts  similarly 
obtained,  the  presence  of  carnosine,  hypoxanthine  and  vitamine  was 
demonstrated  with  certainty.  For  the  purpose  of  vitamine  isolation, 
these  mother  liquors  were  given  by  McLean  to  Cooper  (I.e.  535)  who 
readily  fractioned  out  the  vitamine.  For  the  above  mentioned 
reasons,  it  is  likewise  evident  that  the  growth-promoting  influence 
of  the  lecithides  of  the  liver,  isolated  by  Iscovesco  (612),  was  due  to 
the  presence  of  vitamine.  The  same  is  perhaps  true  for  tethelin, 
isolated  from  the  hypophysis  by  Robertson  (613).  This  lipoid, 
regarded  as  pure,  gave  reactions  for  purine  bases,  as  well  as  a  diazo 
reaction,  so  that  it  is  certain  that  we  are  not  dealing  with  a  pure 

lipoid  in  this  instance. 

228 


RELATIONSHIP   OF   VITAMINES   TO    LIPOIDS  229 

As  regards  vitamine  B,  Cooper  (614),  and  later  Sullivan  and 
Voegtlin  (615),  showed  that  there  is  no  relationship  between  the 
lipoids  and  vitamine  B.  As  for  vitamine  A,  the  behavior  is  quite 
different,  but  still  there  is  no  reason  for  saying  with  Aron  (I.e.  329, 
616)  and  Stepp  (617),  that  lipoids  are  essential  for  life.  In  the  first 
place,  with  a  mixture  consisting  of  the  known  lipods,  lecithin, 
kephalin,  cerebron  and  cholesterol  (Stepp;  I.e.  352),  only  a  slight 
effect  could  be  discerned,  due  perhaps  to  the  cumulative  action  of 
all  the  impurities.  Secondly,  Stepp  (I.e.  351)  showed  that  at  lease 
one '  type  of  animal,  the  pigeon,  could  live  in  the  presence  of  vita- 
mine  B,  without  lipoids.  Recently,  Stepp  (618)  stated  that  dogs 
and  rats  cannot  live  on  a  lipoid-free  diet,  upon  which  the  animal 
develops  paralysis  of  the  hind  extremities,  but  not  beriberi.  An 
addition  of  vitamine  B  prolongs  the  life  of  the  animal  but  cannot 
save  it  from  death.  This  experiment  was  supposed  to  show  that 
animals  die  because  of  a  lack  of  lipoids.  Regarding  the  matter 
from  the  viewpoint  of  the  relationship  of  vitamine  A  to  the  lipoids, 
we  see  that  while  at  first  the  view  was  held  that  this  vitamine  was 
connected  exclusively  with  the  animal  fats,  it  was  found  later  that 
green  leaves  likewise  contain  large  amounts  of  the  same  substance. 
Vitamine  A  is  supposed  to  be  present  in  skimmed  milk,  too,  and 
since  the  latter  is  believed  to  be  free  from  lipoids,  the  relationship 
between  vitamine  A  and  lipoids  must  necessarily  be  non-existent. 
In  particular  the  question  as  to  the  solubility  of  vitamine  A  in  water 
has  not  yet  been  cleared  up;  besides,  in  the  extraction  of  a  fat  with 
water,  as  we  have  already  observed,  the  possible  oxidation  is  to  be 
taken  into  account.  Myers  and  Voegtlin  (I.e.  503)  shook  out  the 
vitamine  of  yeast  with  olive  oil,  and  showed  that  besides  the  vitamine, 
other  nitrogenous  substances  are  taken  up  in  the  oil.  In  this  case, 
we  must  regard  the  total  oil  soluble  portion  as  lipoid  or,  at  any  rate, 
as  fat-soluble.  Since  it  has  never  been  shown  that  vitamine  A  is%a 
constituent  of  a  pure  lipoid,  and  since  up  to  the  present  all  purely 
isolated  lipoids  have  given  negative  results  when  tested  for  their 
activity,  there  is  no  reason  for  considering  vitamine  A  as  being 
associated  with  lipoids;  it  may  ultimately  be  shown  that  this  associa- 
tion is  only  incidental.  Cramer  (619)  described  a  new  type  of 
glandular  fatty  tissue,  which  he  called  "lipoid  gland"  or  "cholesterol 
gland."  This  tissue,  which  is  very  rich  in  cholesterol  and  other 
lipoids,  loses  these  constituents  when  the  animal  is  placed  on  a 


230  THE    VITAMINES 

vitamine-free  diet.  Cramer  believed  that  this  tissue  plays  a  part 
in  the  development  of  avitaminoses,  in  that  it  functions  as  a  vitamine 
reserve  of  the  body.  The  exact  functions  of  this  new  fatty  tissue 
are  not  yet  known,  but  Cramer  thinks  that  a  close  relationship 
exists  between  this  tissue,  the  thyroid  and  the  suprarenals. 


CHAPTER  IV 
THE  ANTISCORBUTIC  VITAMINE — VITAMINE  C 

THE    CHEMISTRY   AND   THE   NATURE    OF   VITAMINE    C 

We  owe  to  Hoist  and  Frolich  (I.e.  359)  our  first  knowledge  on  this 
subject.  They  showed  that  fresh  potatoes,  cabbage,  dandelion, 
carrots,  raspberry .  juice,  lemon  and  sour  dock  juice  contain  this 
vitamine.  They  observed  the  interesting  fact  that  the  various  anti- 
scorbutics differ  markedly  in  the  stability  of  the  vitamine  present, 
evident  on  heating,  storage  and  drying.  As  to  the  reason  for  this, 
we  are  still  in  the  dark.  We  do  not  know  if  in  these  cases  we  are 
dealing  with  different  or  with  the  same  substances,  and  yet  the  varied 
behavior  towards  external  factors  is  not  necessarily  due  to  the  pres- 
ence of  distinct  substances.  The  different  combinations  of  the 
vitamine  and  the  characteristics  of  the  juice  may  be  responsible  for 
this  behavior. 

It  may  be  said  in  general  that  antiscorbutics  exhibit  less  activity 
when  boiled  than  when  in  the  natural  condition;  furthermore,  heating 
to  110°  or  120°C.  is  harmful,  even  more  so  than  boiling  at  100°C. 
It  is  not  only  the  heating  that  exerts  a  harmful  effect,  but  also  the 
incidental  drying,  which  is  in  accord  with  the  experience  gained  in 
the  study  of  human  scurvy.  According  to  Hoist  and  Frolich,  the 
relative  moisture  of  the  drying  chamber  plays  a  part,  since  the  anti- 
scorbutic activity  of  potatoes,  carrots,  dandelion  and  cabbage  is 
more  quickly  lost  at  room  temperature  than  in  an  incubator  at  37°C. 
The  dried  dandelion,  unlike  cabbage,  is  completely  inactivated. 
Vegetable  juices  behave  differently;  for  example,  cabbage  juice  loses 
its  activity  when  heated  for  10  minutes  at  60°,  70°  or  100°C.; 
similarly,  when  it  is  stored  at  ordinary  temperatures  (in  the  presence 
of  preservatives)  or  in  the  ice-box.  In  the  next  chapter  we  shall 
speak  of  the  effect  of  the  cooking  on  the  antiscorbutic  properties  of 
various  foodstuffs. 

Lemon  juice  behaves  quite  differently  from  the  above  mentioned 
substances,  as  does  also  milk.  The  former  may  be  heated  to  110°C. 
for  1  hour  without  any  noticeable  loss  of  activity.  Now  this  juice 
contains  7  per  cent  citric  acid,  and  Hoist  and  Frolich  held  that 

231 


232  THE   VITAMINES 

vitamine  C  was  more  stable  because  of  the  acid  present.  This 
idea  was  confirmed  by  the  fact  that  other  acid  juices,  like  raspberry 
and  sour  dock  juice,  show  the  same  stability  towards  heating.  The 
heat  stability  of  cabbage  and  dandelion  juice  was  markedly  increased 
on  the  addition  of  acid,  but  permanent  stability  could  not  be  obtained. 

The  last  finding  was  made  use  of  in  extractions.  If  freshly  dried 
cabbage  is  extracted  with  alcohol  to  which  a  little  citric  acid  is  added, 
instead  of  with  pure  alcohol,  the  extracts  thus  obtained  are  much 
more  active  on  guinea  pigs.  Other  isolation  experiments,  such  as 
dialysis  or  extraction  with  petroleum  ether,  were  valueless.  Hoist 
and  Frolich  were  unable  to  favorably  influence  scurvy  in  guinea  pigs 
by  intraperitoneal  injections. 

In  a  later  communication,  Hoist  and  Frolich  (620)  stated  that 
cabbage  dried  at  37°C.  and  stored  in  a  desiccator  still  contained 
vitamine  C  after  13  months.  It  was  possible  also  to  obtain  an 
active  substance  by  extracting  cabbage  with  a  mixture  of  alcohol 
and  glycerine.  This  was  also  the  case  with  extractions  made  with 
dilute  alcohol,  to  which  0.5  per  cent  citric  acid  had  been  added.  Hot 
80  per  cent  alcohol  with  0.5  per  cent  citric  acid  likewise  extracts 
vitamine  C  from  freshly  dried  cabbage.  Since  the  experiments  of 
Hoist  and  Frolich  in  1913,  no  real  progress  has  been  made,  in  spite 
of  the  numerous  publications  that  have  appeared. 

Shortly  thereafter,  the  author  undertook  a  chemical  fractionation 
of  vitamine  C,  using  the  same  methods  as  in  the  yeast  fractionation, 
but  the  results  obtained  were  not  very  satisfactory.  The  investiga- 
tion failed  chiefly  because  of  the  lack  of  knowledge  of  scurvy  in 
guinea  pigs,  as  well  as  of  the  behavior  of  vitamine  C  which  at  that 
time  had  hardly  been  recognized.  Although  more  firmly  established 
data  are  available  on  this  subject  at  present,  the  chemistry  of  this 
vitamine  has  thus  far  not  progressed.  In  1912  we  investigated,  in 
this  respect,  milk,  potatoes  and  lime  juice,  using  guinea  pigs  and 
rabbits  kept  on  water  and  oats.  The  guinea  pigs  died  in  about  20 
to  30  days  with  marked  scorbutic  symptoms.  After  the  addition  of 
fresh  potato  juice,  the  animals  died  after  36  days,  while  after  the 
purification  of  this  juice  with  lead  acetate,  the  animals  died  after  27 
days.  A  daily  addition  of  25  cc.  milk  had  a  definite  inhibitory  effect 
on  the  scorbutic  symptoms,  while  on  addition  of  50  cc.  milk,  the 
symptoms  of  scurvy  were  absent  even  after  50  days,  the  animals 
sometimes  gaining  weight ;  we  shall  discuss  the  dissimilar  results  of 


THE   ANTISCORBUTIC   VITAMINE  233 

Chick  and  Hume  in  the  chapter  on  milk.  Protein-free  milk,  prepared 
by  precipitation  of  casein  with  acetic  acid  at  50°C.,  lactalbumin 
removed  by  colloidal  iron  solution,  and  concentrated  by  freezing 
(this  may  perhaps  be  a  practical  method  of  concentrating  vitamine 
C),  exhibited,  in  amounts  corresponding  to  200  cc.  of  fresh  milk, 
only  a  very  slight  activity.  An  alcoholic  extract  of  milk  dried  in 
vacuum  by  us,  possessed  no  activity  in  large  doses,  while  the  residue, 
given  in  doses  of  30  grams  showed  definite  activity  in  some  cases. 
We  then  turned  our  attention  to  lime  juice  (I.e.  101)  which  was 
universally  known  as  a  splendid  antiscorbutic.  For  this  reason  we 
were  unable  at  that  time,  to  explain  why  it  was  inactive  in  guinea 
pigs.  The  same  result  was  also  obtained  with  the  decomposed 
phosphotungstate,  prepared  from  lime  juice.  We  were  beginning  to 
doubt  the  identity  of  scurvy  in  guinea  pigs  with  that  in  man,  though 
recently  the  situation  was  completely  explained  by  Alice  Henderson 
Smith  (621).  In  a  historical  sketch,  she  showed  that  the  statements 
as  to  the  protective  action  of  lime  juice  on  scurvy  were  not  true  for 
the  sour  limes  of  the  West  Indies  used  to-day,  but  were  so  for  sweet 
limes  and  lemons  growing  in  the  Mediterranean  regions. 

The  lime  juice  used  to-day  was  shown  to  be  inactive,  but  we 
demonstrated  the  presence  therein  of  vitamine  B.  Aside  from  this, 
we  isolated  a  terpene  having  the  formula  CisH^Oa,  a  purine  base, 
C6H702N5  (m.p.  282°C.)  and  a  substance  from  the  histidine  fraction, 
melting  at  188°  to  189°C.  and  having  the  formula  C9Hi806N2.  From 
the  choline  fraction,  a  base  was  isolated  as  the  double  platinum  salt, 
which  melted  at  220°C.  and  which  was  given  the  formula,  CsH^N, 
though  a  later  investigation  showed  that  the  substance  was  identical 
with  stachydrine  C7Hi302N.  These  data  are  quoted  here  because 
it  may  later  be  shown  that  one  of  these  substances  is  a  cleavage 
product  of  vitamine  C. 

The  experiments  of  Hoist  and  Frolich  on  the  extractability  of 
vitamine  C  with  alcohol,  were  later  confirmed  by  Freise  (622)  on 
turnips,  as  well  as  by  Freudenberg  (623) .  Subsequently,  Harden  and 
Zilva  (624)  showed  that  when  orange  juice  is  freed  from  citric  acid 
by  precipitation  with  calcium  carbonate,  and  the  mixture  treated 
with  an  equal  volume  of  alcohol,  filtered,  and  the  filtrate  concentrated 
in  vacuum  at  37°C.,  an  active  solution  is  obtained  which,  however, 
loses  its  activity  in  time.  If  lemon  juice  is  treated  similarly,  and 
concentrated  in  acid  reaction,  an  active  residue  is  also  obtained. 


234  THE   VITAMINES 

Later,  Harden  and  Robison  (625)  showed  that  vitamine  C  is  not 
volatile  on  distillation,  that  the  dry  residue  has  almost  the  same 
activity  as  the  original  juice,  and  that  it  may  be  kept  in  a  dry  con- 
tainer for  six  months.  Apple  juices  were  found  to  be  much  poorer 
in  vitamine  C.  This  applies  to  juices  concentrated  in  the  labo- 
ratory; when  prepared  in  large  amounts,  these  preparations  lose  63 
per  cent  of  their  activity,  according  to  Harden  and  Robison  (626). 
Vedder  (626a)  prepared  active  extracts  by  successive  extractions 
with  absolute  alcohol  acetone  and  ethyl  acetate.  Vitamine  C  was 
destroyed  by  phosphotungstic  acid  and  the  last  extract  exhibited 
about  one-seventh  of  the  initial  activity  and  contained  only  traces 
of  nitrogen.  This  led  Vedder  to  the  somewhat  premature  conclusion 
that  vitamine  C  is  nitrogen-free.  Without  going  further,  we  see 
that  active  preparations  of  vitamine  C  may  be  obtained  also  in  the 
absence  of  citric  acid,  and  we  are  therefore  not  yet  certain  whether 
these  acids  possess  protective  qualities. 

Givens  and  McClugage  (627)  dried  orange  juice  in  two  different 
ways.  First,  in  shallow  dishes  at  55°  to  60°C.  for  50  hours;  second, 
according  to  a  method  used  in  drying  milk  (Just-Hatmaker  process), 
in  which  a  quick  drying  at  75°  to  80°C.  is  obtained.  Whereas  the 
second  preparation  was  almost  as  active  as  the  original  juice,  the 
first  lost  a  great  part  of  its  effectiveness.  The  active  products  were 
kept  for  3J  months  without  loss  of  activity.  Stable  preparations 
of  vitamine  C  were  made  by  Dubin  and  Lewi  (628)  and  also  by 
Bassett-Smith  (629).  As  regards  the  stability  of  vitamine  C  to 
alkalis,  a  number  of  papers  have  been  published.  Harden  and  Zilva 
(630)  made  orange  juice  alkaline  with  a  -£$  sodium  hydroxide  and 
kept  this  for  24  hours,  resulting  in  a  loss  of  activity.  Hess  and 
Unger  (631)  carried  out  similar  experiments  and  believed  that,  for 
the  most  part,  storage  in  alkaline  medium  exerted  an  unfavorable 
influence  on  the  activity.  Accordingly,  if  the  juice  is  boiled  up  in  a 
slightly  alkaline  solution  and  immediately  neutralized,  the  activity 
is  only  slightly  diminished.  McClendon  and  Sharp  (632)  investi- 
gated the  reaction  of  natural  juices  to  determine  if  they  are  not  some- 
what alkaline,  which  would  explain  the  destruction  of  vitamine  C 
on  boiling;  however,  most  of  the  juices  were  found  slightly  acid. 

Sommer  and  Hart  (633)  stated  that  the  destructive  action  of  boiling 
on  vitamine  C  was  not  due  to  the  precipitation  of  citrates,  while 
Faber  (634)  found  that  an  addition  of  sodium  citrate  (even  in  a  con- 
centration of  0.25  per  cent)  has  a  destructive  action  on  this  vitamine. 


THE   ANTISCORBUTIC   VITAMINE  235 

As  regards  the  oxidizability  of  vitamine  C,  Fowler  (635)  showed 
that  milk  preserved  with  hydrogen  peroxide  is  scurvy  producing. 
Hess  (636)  believed  that  milk  or  canned  tomatoes  shaken  with  air 
lose  considerably  in  their  antiscorbutic  activity. 

According  to  Zilva  (I.e.  513)  ultra-violet  light  has  no  influence  on 
the  activity  of  vitamine  C.  Adsorption  likewise  does  not  affect  this 
vitamine.  It  may  be  treated  with  fullers  earth  or  colloidal  iron, 
according  to  Harden  and  Zilva  (I.e.  89),  and  it  may  also  be  filtered 
through  a  Berkefeld  filter  without  loss  of  activity.  Zilva  and 
Miura  (636a)  were  unable  to  separate  vitamines  B  and  C  by  means 
of  differential  dialysis. 

As  for  the  significance  of  vitamine  C  in  the  organism,  very  little  is 
known,  and  what  little  knowledge  there  is  available  will  be  described 
under  human  scurvy.  The  question  as  to  whether  the  parenteral 
administration  of  vitamine  C  is  effective  has  been  answered  differently 
by  different  investigators.  Hoist  and  Frolich  (I.e.  359)  gave  this 
vitamine  intraperitoneally,  and  Harden  and  Zilva  (I.e.  624)  found 
it  inactive  on  subcutaneous  injection.  Hess  and  linger  (637),  on 
the  contrary,  working  with  children,  showed  that  intravenous 
therapy  with  weakly  alkaline  orange  juice  is  possible,  and  they 
recommend  this  procedure. 

Here,  too,  as  with  vitamine  B,  Hess  (638)  accepted  first  that  a 
large  addition  of  carbohydrates  hastens  the  onset  of  scurvy,  but 
he  (639)  has  changed  his  view  recently.  Hess  has  tried  by  addition 
of  3  per  cent  flour  to  the  milk  to  hasten  the  onset  of  scurvy  in  chil- 
dren, with  negative  results.  We  do  not  believe,  however,  that  this 
amount  of  carbohydrate  is  sufficient  to  influence  the  effect  one  way 
or  the  other. 


CHAPTER  V 

VITAMINS  CONTENT  OF  VARIOUS  FOODSTUFFS,  IN  THE  NATURAL 
AND  PREPARED  CONDITIONS 

We  have  already  shown  that  during  the  last  few  years  most  food- 
stuffs have  been  investigated  for  their  nutritive  value  and  vitamine 
content.  The  results  obtained  give  us  an  insight  into  the  compara- 
tive values  of  the  above,  and  yet  we  must  be  careful  in  interpreting 
the  data.  First  of  all,  it  should  not  be  forgotten,  as  frequently 
happens  nowadays,  that  the  results  obtained  with  one  animal  species 
cannot,  ipso  facto,  be  applied  to  others. 

For  instance,  it  seems  to  us  unjustifiable  to  determine  the  nutritive 
value  of  an  animal  diet  by  using  animals  that  normally  live  on  plant- 
diet;  this  may  be  the  cause  of  the  wide  differences  between  experi- 
ments on  man  and  those  on  animals.  Variations  in  the  antiscorbutic 
value  of  meat  in  man  and  guinea  pigs  may  be  due  to  just  such 
procedures. 

Another  difficulty  in  the  interpretation  of  results  is  the  lack  of 
quantitative  methods  for  the  estimation  of  the  vitamine;  this  is  most 
apparent  in  the  comparison  of  the  results  obtained  at  the  outset  of 
vitamine  research,  with  those  of  modern  investigations  carried  out 
with  a  better  understanding  of  this  new  subject.  Many  of  the  older 
results  were  obtained  without  paying  any  attention  to  quantitative 
relationships,  and  Chick  and  Hume  (640)  correctly  noted  that  in 
this  regard,  we  should  be  constrained  to  work  quantitatively.  For 
example,  if  we  would  wish  to  study  the  effect  of  alkali  on  vitamines,  it 
does  not  suffice  merely  to  add  alkali  to  the  foodstuff  and  feed  it  to 
the  animal.  We  must  first  determine  the  minimal  protective  dose  of 
the  natural  preparation,  for  if  too  great  a  dose  is  used  from  the  start, 
the  preparation,  in  spite  of  an  extensive  destruction,  may  still  contain 
sufficient  vitamine  to  act  therapeutically,  and  we  would  make  the 
erroneous  conclusion  that  the  treatment  to  which  the  foodstuffs  had 
been  subjected  was  without  influence  on  the  vitamine  content. 

Most  .of  the  animal  experiments  conducted  up  to  the  present 
suffered  because  of  the  fact  that  with  the  addition  of  vitamine  in  the 
form  of  food,  or  in  the  form  of  extracts,  the  composition  of  the  basal 
diet  was  at  the  same  time  altered,  without  having  the  corresponding 

236 


VITAMINE   CONTENT   OF   FOODSTUFFS 


237 


controls  with  no  vitamine  addition.  Most  of  the  investigators  also 
regarded  the  vitamine  content  of  any  particular  diet  as  an  unchange- 
able constant,  as  a  mathematical  certainty.  We  shall  see  in  this 
chapter  that  this  is  not  at  all  the  case,  and  it  will  become  clear  how 
the  various  investigators  could  have  obtained  dissimilar  results  with 
the  same  food. 

After  pointing  out  the  possible  relationship  between  vitamines  and 
appetite  (Mendel  641),  and  the  necessity  of  new  legislation  for  food- 
stuffs with  regard  to  the  vitamines  (Ranwez  642),  we  shall  take  up 
the  description  of  the  treatment  of  foodstuffs  as  related  to  the  vita- 
mine  content.  Here,  there  are  the  effect  of  heating,  cooking,  drying, 
storing,  sterilizing,  and  preserving.  Here,  too,  we  shall  describe  the 
more  important  foodstuffs  commonly  used  in  human  nutrition.  It 
will  be  impossible,  however,  to  go  into  all  of  the  investigations  made 
in  this  respect,  and  for  this  reason  we  shall  put  all  of  the  data  con- 
sidered of  importance  in  the  form  of  a  table  at  the  end  of  this  chapter, 
showing  the  relative  richness  of  the  foodstuffs  in  the  three  known 
vitamines.  The  first  chart  of  this  kind  was  made  by  Cooper  (643) 
in  1912-14,  including  but  a  few  foodstuffs  and  concerning  itself  only 
with  the  vitamine  B  content.  A  similar  table  was  given  in  the 
Report  of  the  Medical  Research  Committee  (I.e.  333)  wherein  it  was 
attempted  to  record  quantitatively  the  values  found.  The  values  for 
vitamine  B  are  recorded,  compared  to  100  for  wheat  germ. 


FOODSTUFFS 

VALUE 

MOISTURE  CONTENT 

Wheat  germ 

100 

per  cent 

10-13 

Wheat  bran  

25 

10-13 

Rice  germ  

200 

10-13 

Pressed  yeast 

60 

70 

Dried  peas  

40 

12 

Lentils  .   . 

80 

EKE  volk 

50 

70 

Ox  liver  

50 

70 

Ox  muscle 

11 

75 

Potatoes  

4  3 

80 

In  the  same  report,  we  see  also  the  first  investigation  of  the  con- 
tent of  other  vitamines.  All  of  these  values  were  obtained  on 
animals  and  apply  strictly  only  to  the  species  upon  which  the  results 


.238  THE   VITAMINES 

were  determined.  This  is  true  especially  as  regards  quantitative 
relationships.  Chick  and  Dalyell  (644)  prepared  another  table  with 
regard  to  vitamine  C,  where  the  values  are  based  on  100  for  lemon 
juice. 

Fresh  orange  juice  or  cabbage  leaves 110 

Fresh  raw  orange  juice 100 

Beet  juice  (swede) 60 

Green  beans 30 

Sprouted  fresh  peas 30 

Carrot  juice 7.5 

Red  beet  juice 7.5 

Meat  juice  (ox) 7.5 

Potato  (cooked  39  minutes) 7.5 

Fresh  cow's  milk 1-1 . 5 

In  our  table,  we  were  not  satisfied  to  give  only  the  vitamine  value, 
but  also  the  nutritive  value  of  the  proteins  of  various  origin.  This 
rubric  will,  perhaps,  help  us  to  demonstrate  the  difference  between 
the  nutritive  value  of  plant  and  animal  protein,  which  may  be  of 
importance  in  relation  to  the  etiology  of  pellagra  and  war  edema; 
unfortunately,  however,  the  data  found  on  this  subject  are  very 
meagre.  It  appears  to  us  not  impossible  that  some  protein  substances 
may  contain  adsorbed  either  a  fourth  vitamine,  or  a  hitherto  unknown 
amino  acid,  essential  to  life. 

INFLUENCE   OF  HEATING  AND   COOKING   ON   THE   VITAMINE    CONTENT1 

We  owe  to  Schiiffner  and  Kuenen  (I.e.  56)  the  first  data  on  this 
subject  of  such  practical  importance.  Studies  on  the  stability  of 
vitamine  B  in  foodstuffs  were  undertaken  also  by  Grijns  (I.e.  52). 
Recently  this  field  was  systematically  investigated  by  Chick  and  Hume 
(645),  using  an  improved  technique.  They  studied  the  influence  of 
heat  on  vitamine  B  of  wheat  germ  and  found  that  a  temperature 
of  100°C.  for  two  hours  has  little  influence,  while  a  temperature  over 
120°C.  is  quickly  followed  by  destruction.  Miller  (646)  found  that 
cooking  and  autoclaving  (at  115°C.  for  45  minutes)  had  no  effect  on 
carrots  and  beans;  this  was  confirmed  by  Whipple  (647)  for  cab- 
bage and  onions.  In  the  last  two  investigations  the  content  of 
vitamine  B  was  determined  by  the  yeast  method  already  described 

1  A  good  review  of  the  known  data  is  found  in  the  paper  by  Emmett  and 
Luros  (I.e.  94). 


VITAMINE    CONTENT   OF   FOODSTUFFS  239 

by  us,  but  was  not  controlled  by  animal  experiments.  The  stability 
of  vitamine  C  against  heat  depends  very  much  on  the  nature  of  its 
source.  Weill  and  Mouriquand  (648)  stated  that  this  vitamine  is 
very  labile,  but  noted  that  lemon  juice  may  be  heated  to  110°C.  for  1 
hour  without  marked  loss  in  activity.  Delf  and  Tozer  (649),  co- 
workers  of  Chick,  carried  out  investigations  on  cabbage:  heating  for 
1  hour  at  60°C.  decreased  the  antiscorbutic  value  about  70  per  cent, 
at  90°C,  about  90  per  cent,  while  on  short  heating  (20  minutes),  at 
90°  or  100°C.,  the  activity  is  decreased  only  about  70  per  cent;  we 
see  from  this  that  the  chief  influence  is  produced  by  the  time  of  heat- 
ing and  not  by  the  temperature.  Strudwick  (650)  investigated,  in 
this  respect,  the  influence  of  cooking  in  a  fireless  cooker  in  which  the 
time  of  cooking  was  markedly  prolonged.  The  results  showed  that 
a  rapid  heating  is  much  superior  to  the  above  method. 

Delf  (65 1)2  investigated,  in  the  same  way,  the  relationship  between 
cabbage  leaves  and  cabbage  juice,  showing  that  in  the  natural  con- 
dition, vitamine  C  is  much  more  resistant.  Turnip  juice  (swede) 
was  shown  to  be  far  more  stable  than  cabbage  juice.  Orange  juice 
heated  to  130°C.  lost  50  per  cent  of  its  activity.  Givens  and  McClug- 
age  (652)  found  tomatoes  quite  resistant  to  heat.  However,  if  they 
are  heated  for  15  minutes  or  more  at  100°C,  a  greater  amount,  in 
comparison  with  the  crude  material,  must  be  fed  in  order  to  protect 
guinea  pigs  against  scurvy.  Aside  from  heating,  oxidation  appar- 
ently also  plays  a  role  in  the  inactivation  of  vitamine  C.  Rossi  (653) 
fed  guinea  pigs  on  oats  and  hay  sterilized  in  open  kettles  at  126°C., 
with  the  result  that  the  animals  died  of  scurvy.  However,  a  different 
result  was  obtained  when  the  food  was  heated  as  above  in  closed 
kettles;  on  this  food,  the  guinea  pigs  were  still  alive  after  two  months. 
Delf  (I.e.  651)  also  believed  that  in  the  presence  of  air  the  inactiva- 
tion of  vitamine  C  was  hastened,  and  this  condition,  according  to 
Steenbock,  Boutwell  and  Kent  (I.e.  594),  and  others,  plays  a  part 
in  the  destruction  of  vitamine  A. 

The  destruction  of  vitamines  by  heat  depends  on  still  another 
factor — the  chemical  reaction  of  the  starting  material.  In  this 
connection,  the  data  are  not  so  certain  for  vitamine  B.  While  McCol- 
lum  and  Simmonds  (I.e.  315)  reported  on  the  supposed  destruction 

2  In  comparing  the  temperature  coefficient  of  the  heat  destruction  of 
vitamine  C  with  toxins  and  ferments,  Delf  concluded  that  this  vitamine 
must  possess  a  relatively  simple  chemical  structure. 


240  THE    VITAMINES 

of  this  vitamine  by  alkalis,  Daniels  and  McClurg  (654)  stated  that 
they  observed  no  destr  iction  of  cabbage  and  soya  beans  so  treated. 
It  is  possible,  however,  that  the  latter  investigators  fed  too  much 
of  the  active  material.  According  to  Daniels  and  Heisig  (655),  the 
effect  of  the  addition  of  soda  upon  vitamine  C  was  deleterious.  In 
this  connection,  Hess  and  Unger  (656)  point  out  that  soda  is  used  in 
the  preparation  of  proprietary  infant  foods.  This  is  true  also  of 
commercial  cod  liver  oil  emulsions,  which  give  an  alkaline  reaction, 
and  to  which  soda  is  added  in  order  to  obtain  a  better  emulsion.  Yet 
we  have  no  exact  experiments  to  show  whether  the  activity  of  the 
above  emulsions  is  affected  by  the  manner  of  preparation.  Experi- 
ments started  by  us  have  not  yielded  any  definite  results  up  to  the 
present.  We  must  say,  however,  that  the  vitamine  A  of  cod  liver 
oil  must  be  quite  resistant  in  order  to  withstand  the  treatment 
undergone  during  the  process  of  manufacture. 

It  has  often  been  stated  that  the  stability  of  orange  juice  is 
explained  by  the  acid  reaction  of  the  medium.  Still,  according  to 
the  findings  of  Delf  (I.e.  651),  the  neutralization  of  the  juice  did  not 
have  any  harmful  effect. 

Summing  up  the  subject  matter  of  this  important  chapter,  we  may 
say  that  the  stability  of  the  vitamines  on  heating  depends  not  on 
the  temperature  but  on  the  duration  of  the  heating.  In  this  respect, 
the  view  of  Givens  and  McClugage  (657)  is  of  interest ;  they  believed 
that  when  antiscorbutics  are  heated  at  a  low  temperature  for  a  long 
time,  as  in  gradual  drying,  the  ferments  contained  in  the  tissues 
destroy  the  vitamine.  If  the  ferments  are  inactivated  by  rapid 
heating  at  high  temperatures,  then  vitamine  C  is  much  more  resist- 
ant. These  statements  naturally  require  further  corroboration.3 
Aside  from  this,  the  vitamines  seem  to  be  more  stable  in  the  natural 
condition  than  in  the  form  of  extracts  or  pressed-out  juices.  Oxidation 
plays  a  big  part  in  the  destruction  of  vitamines  A  and  C,  while  the 
chemical  reaction  of  the  substrate  plays  a  part  in  the  destruction 
of  all  the  vitamines. 

3  This  behavior  may  be  brought  into  relationship  with  the  stability  in 
acid  solution,  if  it  is  assumed  that  these  ferments  act  in  alkaline  solution; 
we  have  in  mind  here  the  oxidases,  the  action  of  which  is  delayed  in  acid 
medium. 


VITAMINE    CONTENT   OF   FOODSTUFFS  241 

The  vitamine  content  of  cooking  water 

That  vitamines  may  be  lost  by  discarding  the  cooking  water,  was 
first  stated  by  Schiiffner  and  Kuenen  (I.e.  56).  In  addition,  vita- 
mi  nes  are  lost  in  other  water-soluble  substances  and  it  is  therefore  not 
without  reason  that  Hindhede  (658)  pointed  out  the  great  nutritive 
value  of  soups,  especially  vegetable  soups.  Hill  (659)  stated,  for 
example,  that  on  cooking  potatoes,  losses  ensued  which  represented 
70  per  cent  of  all  water-soluble  substances.  From  the  viewpoint  of 
vitamines,  the  same  thing  was  pointed  out  by  a  number  of  workers — 
Denton  (660),  Daniels  and  McClurg  (I.e.  654),  Miller  (I.e.  646)  and 
Whipple  (I.e.  647).  The  last  two  investigators,  in  particular,  found 
that  by  discarding  the  cooking  water,  30  to  70  per  cent  of  the  sub- 
stance necessary  for  the  growth  of  yeast  was  lost. 

THE   INFLUENCE    OF   DRYING 

Since  the  question  of  the  dehydration  of  foodstuffs  has  already 
assumed  practical  importance,  it  is  regrettable  that  comparatively 
little  is  known  of  its  effect  on  vitamines.  Murlin  (661)  believed  that 
the  nutritive  value  of  dried  foodstuffs  is  strictly  comparable  with 
that  of  fresh.  Prescott  (662)  put  the  practical  side  of  this  problem 
to  the  fore,  in  relation  to  transportation  and  storage,  particularly  for 
the  provisioning  of  any  army  in  the  field.  He  stated  also  that  the 
loss  of  vitamine  C  on  drying  may  be  prevented  by  modifying  the 
procedure.  Givens  and  Cohen  (663)  investigated  this  question,  using 
cabbage  and  potatoes.  Drying  in  a  stream  of  air  at  40°  to  52°C. 
resulted  in  an  appreciable  loss  of  activity.  With  the  simple  drying, 
the  preparation  of  foods  is  not  yet  completed,  and  practically  two 
other  factors  are  to  be  considered — the  storing  and  the  cooking  just 
before  eating.  These  two  factors  have  unfortunately  not  been 
considered  by  most  investigators;  nevertheless,  it  was  found  that 
when  cabbage  or  potatoes  were  first  heated  or  cooked  and  then  dried, 
no  more  vitamine  C  was  left,  as  shown  in  guinea  pig  experiments.4 

4  Hoist  and  Frolich  (663a)  found  that  if  moisture  is  excluded  as  much  as 
possible  to  diminish  the  chances  of  hydrolysis,  a  stable  cabbage  preparation, 
as  regards  vitamine  C  content,  may  be  obtained.  The  dehydration  was 
carried  out  in  vacuum  at  37°C.  in  presence  of  phosphorus  pentoxide,  which 
is  replaced  as  soon  as  it  becomes  moist.  This  preparation  was  kept  for  10,  18, 
and  26  months  and  found  active. 


242  THE   VITAMINES 

Givens  and  McClugage  (664)  found  tomatoes  more  resistant  in  this 
respect,  at  least  in  relation  to  storage  after  drying,  since  after  three 
months,  the  activity  was  still  retained.  Subsequent  cooking  experi- 
ments appear  to  have  not  been  made,  but  are  not  particularly  impor- 
tant inasmuch  as  tomatoes  may  also  be  used  raw.  Shorten  and  Ray 
(665)  dried  various  kinds  of  vegetables  in  the  sun,  and  investigated 
their  vitamine  B  and  C  content.  The  results  showed  that  vitamine 
C  of  carrots,  onions,  and  cabbage  were  still  present  in  sufficient 
amounts  for  guinea  pigs.  Spinach,  on  the  contrary,  lost  this  prop- 
erty. We  find  no  statements  here  to  the  effect  of  ageing  and  subse- 
quent cooking.  Falk,  McGuire  and  Blount  (666)  studied  the  ferments 
of  dehydrated  vegetables.  They  found  them  inactivated  and  pointed 
out  the  similarity  to  the  resistance  of  the  vitamines.  Givens  and 
McClugage  (I.e.  657)  published  a  very  careful  investigation  upon  the 
drying  of  potatoes.  Taking  their  work  as  a  whole,  we  find  that  when 
potatoes  undergo  a  series  of  treatments,  the  individual  effects  are 
summed  up,  resulting  in  a  totally  inactive  product.  If  the  potatoes 
are  boiled,  steamed  or  baked  and  then  dried,  an  inactive  product 
is  obtained,  as  a  rule.  Drying  alone  does  not  have  this  destructive 
effect.  The  vitamine  was  least  affected  when  the  potato  was  baked 
in  the  jacket  (perhaps  on  account  of  absence  of  air)  and  then  dried. 
Addition  of  acids  before  treatment  did  not  have  the  desired  effect. 
Potatoes  dried  in  vacuum  also  lost  considerable  activity.  We  have 
already  seen  that  quick  drying  at  high  temperatures  is  less  destructive 
than  gradual  drying  at  low  temperatures.  Ageing  and  subsequent 
drying  were  not  investigated,  although  it  is  clear  from  what  has 
already  been  said  that  vitamine  C  of  potatoes  is  not  resistant. 

CANNING 

This  subject  too  has  not  yet  been  extensively  investigated.  We 
believe  that  for  this  question,  the  foodstuffs  may  be  classified  in  two 
groups — those  used  raw,  and  those  to  be  cooked  before  consumption. 
We  find  the  conditions  here  similar  to  those  in  drying.  Bigelow  (667) 
described  the  problems  arising  in  canning.  Different  sources  of 
supply  give  rise  to  different  conditions  so  that  general  conclusions 
can  hardly  be  drawn  as  yet.  Hess  and  linger  (668)  found  that  canned 
tomatoes  still  showed  a  splendid  antiscorbutic  action  in  children,  and 
adopted  this  antiscorbutic  in  their  children  practice;  it  is  specially 
suitable  for  institutions  since  the  expensive  orange  juice  may  be 


VITAMINE    CONTENT   OF   FOODSTUFFS  243 

substituted  by  something  cheaper.  This  finding  is  of  practical 
significance,  since  in  the  anticipation  of  scurvy  epidemics,  canned 
tomatoes,  which  appear  to  keep  well,  may  be  used,  though  it  should 
be  remembered  (Hess  and  linger,  I.e.  631)  that  the  activity  decreases 
on  cooking.  Daniels  and  McClurg  (I.e.  654)  believe  that  the  canning 
proces  does  not  destroy  vitamine  B;  it  seems  to  us  that  an  absolute 
proof  of  this  has  not  been  obtained  in  canning,  although  they  admit 
that  changes  initiated  during  canning  may  progress  automatically  dur- 
ing storage.  Campbell  and  Chick  (669)  prepared  cabbage  and  beans 
in  a  manner  similar  to  that  used  in  canning,  and  found  that  these 
substances  lost  about  66  per  cent  and  75  to  90  per  cent  of  their 
activity,  respectively.  Vitamines  A  and  B  were  also  affected  although 
the  investigators  explained  it  by  losses  of  vitamine  A  into  the  cook- 
ing water.  When  this  was  given  to  guinea  pigs,  it  resulted  in  better 
growth.  We  must  assume,  therefore,  with  Campbell  and  Chick, 
that  vitamine  A  went  over  into  the  cooking  water. 

AGEING   AND   STORING 

The  last  factors  to  be  taken  up  in  this  connection  are  those  of  ageing 
and  storage.  Hulshoff  Pol  (670)  could  cure  human  beriberi  with 
puree  of  peas,  although  peas,  which  were  three  years  old,  were  entirely 
inactive.  Weill,  Mouriquand  and  Michel  (I.e.  392)  found  the  same 
to  be  true  of  denatured  meat,  stored  for  14  months.  For  cats,  this 
meat  was  much  more  harmful  than  freshly  denatured  meat.  Delf 
and  Skelton  (671)  dried  cabbage  and  stored  it  at  ordinary  tem- 
perature; after  2  to  3  weeks,  the  preparation  lost  93  per  cent  of  its 
activity  and  after  3  months,  96  to  97  per  cent.  Hess  and  Unger  (672) 
found  no  vitamine  C  in  prunes,  while  orange  peel  was  active  even 
after  3  months.  The  storage  of  orange  juice  has  a  definite  influence, 
since  in  old  juice,  only  half  of  the  activity  remained.  Dried  carrots, 
kept  for  3  to  5  weeks  and  for  7  months,  were  quite  inactive.  Hess 
and  Unger  concluded,  from  this  that  dried  vegetables  actually  retain 
their  nutritive  value,  though  as  regards  vitamine  C,  they  cannot 
replace  the  fresh  natural  products.  The  ageing  applies  not  only  to 
storing  and  drying,  but  also  to  fresh  vegetables.  In  this  respect, 
Hess  and  Unger  (I.e.  631)  investigated  carrots  of  various  ages.  While 
young  carrots  retained  their  activity  even  after  cooking  and  drying 
in  a  current  of  air,  and  storing  for  3  months,  this  was  not  the  case 
with  old  carrots. 


244  THE   VITAMINES 

From  what  has  been  said,  we  see  that  universal  rules  as  to  the 
stability  of  the  vitamines  cannot  be  formulated.  Every  single  case 
must  be  investigated  separately,  since  we  do  not  yet  know  of  any 
rule  for  determining  the  stability  or  lability  of  these  substances. 
Evidently,  the  vitamine  content  of  foodstuffs  prepared  for  ingestion 
depends  considerably  upon  its  original  content  in  the  natural  state. 
This  original  content  may  vary  wit'i  the  country  in  which  it  is 
produced,  with  the  condition  of  the  soil,  the  climate  and  so  on. 
This  is  true  to  our  primary  sources  of  vitamines.  As  for  the  second- 
ary sources,  animal  products,  the  question  is  much  more  complicated 
and  depends,  for  the  most  part,  upon  known  factors,  but  partly 
upon  unknown  factors  too,  which  we  have  already  discussed. 

CEREALS 

Under  this  heading  we  shall  describe  not  only  the  cereals,  such  as 
wheat,  barley,  rye  and  maize  but  also  the  breadstuff s.  Although  an 
etiological  relationship  between  the  refining  of  cereals  for  human 
consumption  and  beriberi  has  been  suspected  for  a  long  time,  and  in 
fact  demonstrated,  the  economic  conditions  arising  out  of  war  have 
stimulated  interest  in  the  question  and  in  established  knowledge  on 
the  subject.  In  almost  all  countries,  the  investigators  have  endeav- 
ored to  utilize  the  nutritive  value  of  cereals  to  the  best  possible 
advantage.  To  the  problem  of  the  economic  utilization  of  the  grain 
kernels,  another  factor  is  added  which  we  have  already  noted  in  the 
etiology  of  avian  beriberi,  namely,  that  in  most  cases,  vitamines  A 
and  B  are  localized  in  a  certain  portion  of  the  kernel.  Because  of 
this,  modern  refining  may  result  in  a  more  or  less  complete  removal 
of  these  essential  factors.  It  is  indeed  true  that  under  normal  con- 
ditions, when  the  population  eats  other  foodstuffs,  in  addition  to 
bread,  there  is  no  immediate  danger  of  an  avitaminosis,  and  yet  we 
must  remember  that  at  present  bread  may  constitute  a  large  per- 
centage of  the  diet.  Although  whole  wheat  bread  is  used  almost 
exclusively  in  many  countries,  in  more  cultured  sections,  white  bread 
is  used  in  preference.  Aside  from  the  aesthetic  factor,  some  scientific 
facts  are  known  which  make  it  desirable  to  mill  the  grain.  One  of 
the  reasons  frequently  cited  is  the  fact  that  flour  from  whole  wheat 
keeps  with  greater  difficulty.  In  fact,  the  whole  grain,  very  rich  in 
fat,  is  very  difficult  to  store.  The  polishing  of  rice  is  often  defended 
on  these  grounds,  and  Ottow  (I.e.  518)  proposed  measures  for  increas- 


VITAMINE    CONTENT   OF   FOODSTUFFS  245 

ing  its  conservability.  This  is  even  more  applicable  to  corn,  so  rich 
in  fat.  Winton,  Burnet  and  Bornmann  (673)  state  that  corn  may 
be  stored  much  better  if  the  germ  has  been  removed.  The  second 
reason  why  it  is  difficult  to  introduce  whole  grain  in  practice,  is  that 
many  investigators  are  of  the  opinion  that  bread  baked  with  the 
chaff  in  uneconomic.  The  assimilation  of  this  bread  is  supposed  to 
be  not  only  poor,  but  it  occasions  losses  in  other  foodstuffs  simulta- 
neously consumed.  Still,  the  investigators  are  not  agreed  upon  this 
point.  While  Hindhede  (674),  Rohmann  (675),  Eijkman  (676), 
Hulshoff  Pol  (677),  Butcher  (678)  and  Stoklasa  (679)  believe  that 
whole  wheat  bread  is  very  well  utilized;  we  also  encounter  views, 
such  as  those  of  Pugliese  (680),  van  Leersum  and  Munk  (681)  and 
Rubner  (682)  who  asserted  that  such  breadstuffs  are  poorly  utilized 
and  in  fact  provoke  losses,  as  has  been  noted  above. 

Since  the  time  of  Liebig  (683),  we  have  known  that  milling  of 
grain  decreases  its  nutritive  value;  Liebig  said:  "No  single  food- 
stuff loses  its  value  so  readily  as  whole  grain  through  the  modern 
process  of  milling.  The  whiter  the  flour  the  less  nutritive  value  it 
possesses."  In  1871,  Magendie  (684)  knew  that  pigeons  die  when  fed 
on  fine  wheat  flour,  while  on  rye,  they  do  very  well.  Dogs  on  wheat 
flour  die  after  40  days,  while  on  coarse  "soldiers'  bread"  they  continue 
to  live.  Oseki  (685)  investigated  the  value  of  various  kinds  of  bread 
on  mice.  He  found  rye  and  army  bread  suitable,  while  wheat  bread 
and  barley  flour  were  unsuitable.  Weill  and  Mouriquand  (686) 
chose  to  add  extract  of  bran,  instead  of  the  bran  itself,  in  baking 
bread,  whereupon  the  addition  of  the  occasionally  objectionable 
cellulose  was  obviated.  Sherman,  Rouse,  Allen  and  Woods  (687) 
believed  in  the  greater  nutritive  value  of  the  whole  wheat  kernel  as 
compared  with  white  bread,  judging  from  their  rat  experiments. 

Our  knowledge  as  to  the  localization  of  the  vitamines  in  grain 
kernels  has  undergone  some  modification  in  recent  years.  While 
the  first  workers  in  this  field  stated  that  the  protective  substances 
are  found  in  the  "silver  skin"  (a  thin  membrane  surrounding  the 
rice  kernel),  Fraser  and  Stanton  thought  the  protective  substances 
were  to  be  found  in  the  aleurone  layer.  Later,  McCollum  and  Davis 
(688)  showed  that  vitamine  B  is  found  chiefly  in  the  germ.  Chick 
and  Hume  (689)  found  this  vitamine  in  rice,  wheat  and  corn,  partially 
also  in  the  pericarp.  Voegtlin,  Lake  and  Myers  (690)  believed  that 
they  could  show  by  animal  experiments  that  vitamine  B  of  wheat 


246  THE   VITAMINES 

and  maize  occurs  chiefly  in  the  germ,  as  well  as  in  the  pericarp; 
in  this  they  adopted  the  view  of  Chick  and  Hume  at  first.  Later  how- 
ever, Voegtlin  and  Myers  (I.e.  271)  showed  that  when  wheat  or  corn 
kernels  (see  fig.  53-54)  are  cut  into  two  portions,  and  the  part  without 
the  germ  fed  to  animals,  results  are  obtained  which  indicate  an 
absolute  lack  of  vitamine  B;  therefore,  the  vitamine  must  be  in  the 
germ  only.  However,  it  must  not  be  forgotten  that  by  the  procedure 
followed  by  Voegtlin  and  Myers,  other  constituents  are  removed  (for 
example,  protein),  and  that  possibly  the  vitamine  is  also  present  in 
the  remaining  part  of  the  grain  in  sub-optimal  amounts.  Funk  and 
Dubin  (I.e.  493),  showed  that  the  substance  promoting  the  growth 
of  yeast  was  found  in  the  germ,  but  it  was  likewise  present  outside 
of  the  germ  in  appreciable  quantities.  For  practical  purposes, 
according  to  Greig  and  Curjel  (691),  it  seems  sufficient  when  wheat 
germ  is  added  to  zwieback  to  the  extent  of  15  per  cent.  This  measure 
was  adopted  for  the  English  colonial  army  in  order  to  protect  the 
personnel  against  beriberi. 

When  cereal  flours  are  heated  in  baking,  the  vitamine  B  content  is 
altered.  Weill  and  Mouriquand  (692)  heated  decorticated  barley  to 
120°C.  for  1J  hours.  When  this  was  fed  to  pigeons,  they  died  after 
30  days  with  definite  beriberi  symptoms.  On  white  rice,  this  disease 
appears  in  18  to  25  days  and  on  sterilized  rice,  in  9  to  16  days.  They 
believed,  therefore,  that  decorticated  grain  kernels  still  contained 
vitamine,  especially  since  the  symptoms  appeared  sooner  when  the 
food  was  sterilized.  Chick  and  Hume  (I.e.  645)  heated  wheat  germs 
to  high  temperatures,  whereupon  they  observed  inactivation  at 
120°C.,  but  not  on  heating  at  100°C.  for  2  hours.  From  these  results, 
they  believed  that  baking  does  not  affect  vitamine  B  to  any  great 
extent.  If  yeast  is  used  in  the  dough,  then  a  part  of  the  B  vitamine 
of  the  bread  may  arise  from  this  source. 

We  have  frequently  touched  upon  the  point  that  seeds  develop 
vitamine  C  upon  germination.  This  fact,  first  observed  by  Fiirst, 
was  further  developed  for  practical  purposes  by  Chick  and  Delf 
(693),  and  in  addition  to  cereals,  was  confirmed  in  peas  and  beans. 
Weill  and  Mouriquand  (694)  conducted  similar  experiments  with 
germinating  barley  and  later  they  showed,  in  conjunction  with 
Perronet  (695),  that  vitamine  C  develops  in  greater  amounts  only  in 
the  later  stages  of  the  germination. 


VITAMINE    CONTENT   OF   FOODSTUFFS 


247 


FIG.  52.  1,  UNPOLISHED  RICE;  8,  PARBOILED  RICE;  3,  POLISHED  RICE 
(FRASER  AND  STANTON) 


248  THE    VITAMINES 

Rice 

We  have  already  touched  upon  most  of  the  data  on  rice,  including 
those  of  Fraser  and  Stanton  (I.e.  49),  who  showed  by  means  of 
illustrations  how  the  loss  of  substances  occurs  during  the  polishing  of 
rice.  From  the  appended  illustration,  we  see  why  such  great  differ- 
ences in  the  number  of  beriberi  cases  are  apparent  with  the  use  of 
polished  and  parboiled  rice.  The  question  as  to  whether  polished 
rice  still  contains  vitamine  B  has  likewise  already  been  discussed. 
It  is  important  now  to  determine  whether  this  rice  contains  vitamine 
A,  but  we  find  no  observations  on  this  point.  It  would  be  of  special 
interest  to  see  whether  pigeons  can  be  maintained  for  a  long  time  on 
polished  rice  with  the  addition  of  vitamine  B.  Funk  and  Dubin 
(696)  have  started  experiments  in  which  pigeons  are  being  fed  on 
rice  to  which  has  been  added  5  per  cent  casein  and  3  per  cent  salts, 
and  the  mixture  autoclaved  at  120°C.  for  3  hours.  By  means  of  the 
prolonged  heating,  it  was  expected  that  the  rest  of  the  vitamine  A 
would  be  destroyed.  The  pigeons  have  gained  in  weight  and  have 
remained  in  good  health  for  over  six  months.  Despite  the  fact  that 
others  have  investigated  this  problem,  it  is  not  yet  certain  whether  half 
grown  pigeons  can  live  without  an  addition  of  vitamine  A,  or  whether 
it  must  be  concluded  that  autoclaved,  polished  rice  still  contains  traces 
of  vitamine  A.  As  regards  the  Nutritive  value  of  rice  polishings, 
Mattei  (697)  believed  that  pigeons  could  not  be  maintained  on  rice 
polishings  alone;  he  found  that  an  addition  of  white  rice  was  neces- 
sary. Wise  and  Broomel  (698)  investigated  the  effect  of  the  modern 
milling  methods  on  the  composition  of  the  remaining  rice  kernel.  On 
polishing,  the  husk  germ,  6  layers  of  bran  and  a  part  of  the  seventh 
layer  were  removed,  so  that  the  total  loss  in  weight  was  about  10 
per  cent.  On  analysis,  it  was  possible  to  show  a  loss  of  70  per  cent 
ash,  85  per  cent  fat,  and  10  per  cent  protein.  To  prevent  these 
losses,  Guareschi  (699)  has  suggested  the  use  of  rice  which  is  not  so 
highly  milled. 

Barley 

According  to  the  older  statements  about  barley,  it  seems  that  its 
vitamine  B  is  not  localized  in  the  same  way  as  in  other  cereals.  In 
the  older  Japanese  data,  for  example,  that  of  Saneyoshi  (700),  we  find 
that  barley  can  cure  avian  beriberi,  whereas  Weill  and  Mouriquand 


VITAMINE    CONTENT    OF   FOODSTUFFS 


249 


(701)  used  decorticated  barley  to  produce  beriberi.  Steenbock,  Kent 
and  Gross  (702)  observed  that  for  rats,  barley,  like  other  cereals,  was 
deficient  in  vitamine  A,  protein  and  salts. 

Oats 

McCollum,  Simmonds  and  Pitz  (I.e.  372)  regarded  the  composition 
of  oats  as  unfavorable  for  the  nutrition  of  animals.  Tested  on  rats, 
it  showed  a  lack  of  vitamine  A,  protein  and  salts.  This  applies  much 
less  to  pigeons,  as  we  ourselves  have  shown. 

Wheat 

Voegtlin  and  Myers  (I.e.  271) 
believed  that  vitamine  is  found  ex- 
clusively in  the  germ;  we  have 
already  published  our  findings  in 
this  respect  (I.e.  271).  How  the 
various  parts  of  the  wheat  kernel 
are  related  to  each  other,  we  may 
see  from  the  following  diagram, 
taken  from  the  work  of  the  above 
investigators,  from  which  we  may 
note  the  method  of  sectioning  used 
by  them  in  their  experiment  on 
the  localization  of  vitamine  B. 
McCollum,  Simmonds  and  Pitz 
(703)  investigated,  in  particular, 
the  vitamine  value  of  wheat  germ. 
Qualitatively  considered,  this  por- 
tion of  the  wheat  has  been  shown  to  be  adequate,  but  for  practical 
reasons,  an  addition  of  salts  must  be  given.  Vitamine  B  occurs 
in  the  germ  in  large  quantities;  vitamine  A,  on  the  other  hand, 
in  smaller  amounts.  Osborne  and  Mendel  (704)  investigated  the 
nutritive  value  of  various  milling  products  of  wheat  kernels,  and 
did  not  believe  in  the  exclusive  localization  of  vitamine  B  in  the 
germ.  The  contiguous  portion  was  also  rich  in  vitamine  B,  as  was 
also  the  periphery  of  the  kernel,  the  endosperm  and  bran  not  con- 
taining very  much. 


FIG.  53.  WHEAT  KERNEL  SHOWING 

DIFFERENT  LAYERS  (VOEGTLIN- 

MYERS) 


250  THE    VITAMINES 

Maize 

Because  of  a  possible  etiological  relationship  of  corn  consumption 
to  pellagra,  the  numerous  investigations  on  the  nutritive  value  and 
composition  of  corn  and  corn  products  are  of  special  importance. 
Bezzola  (705)  showed  that  corn  does  not  provide  an  adequate  diet 
for  the  maintenance  of  guinea  pigs  in  good  health;  they  lose  hair, 
develop  severe  diarrhea  and  subsequently  die.  A  difference  between 
the  use  of  good  and  bad  corn  was  not  observed.  Lucksch  (706) 
found  that  guinea  pigs,  fed  on  good  corn,  lost  their  hair,  showed  a 
hyperemia  of  the  intestinal  mucosa  and  an  enlargement  of  the  adre- 
nals. On  a  mixture  of  corn,  flour  and  greens,  they  lost  hair  never- 
theless, and  developed  paralysis  of  the  hind  legs  and  catarrh  of  the 
small  intestine.  Corn  was  likewise  shown  to  be  inadequate  for 
rabbits  and  dogs,  the  results  obtained  in  spring  being  more  definite 
than  those  in  autumn.  The  blood  of  the  experimental  animals  was 
investigated  and  found  sterile  at  all  times,  v.  Neusser  (707) 
reported  on  a  disease  of  horses,  called  "emmaisadura"  in  Mexico  and 
Colombia  supposed  to  be  caused  by  rotted  corn.  The  symptoms  of 
this  disease  are  emaciation,  dizziness,  lethargy,  cramps,  wildness,  loss 
of  hair  and  teeth  and  decay  of  the  hoofs.  Hoist  (708)  remarked  on 
the  frequent  occurrence  of  scorbutic  symptoms  in  pellagra,  especially 
the  porosity  of  the  bones.  He  repeated  Lucksch's  experiments  with 
guinea  pigs  and  came  to  the  conclusion  that  the  disease  described  by 
him  could  be  prevented  by  the  addition  of  fresh  cabbage.  In  par- 
ticular, the  constant  loss  of  hair  could  be  overcome  although  the 
experiments  were  carried  out  in  spring.  The  whole  syndrowe  was 
thereupon  regarded  by  Hoist  as  scurvy.  Baglioni  (709)  likewise 
carried  out  some  feeding  experiments  with  corn  products  on  guinea 
pigs.  In  the  light  of  modern  investigations,  Hoist  was  right  when  he 
characterized  as  scurvy,  the  disease  developed  by  guinea  pigs  fed  on 
corn.  Since  in  most  corn-feeding  experiments,  no  antiscorbutic  was 
given,  the  resulting  condition  was  complicated  by  scorbutic  symptoms 
to  such  an  extent,  that  these  investigations  are  of  no  value  in  the 
explanation  of  pellagra.  The  problem  appears  in  a  different  light 
when  birds  are  used  as  experimental  animals.  Ohler  (I.e.  262) 
observed  that  when  decorticated  maize  is  fed  to  chickens,  they 


VITAMINE    CONTENT   OF   FOODSTUFFS  251 

develop  beriberi.  Under  the  same  experimental  conditions,  Driscoll 
(710)  noted  a  condition  which  he  thought  was  similar  to  pellagra  in 
man;  among  other  symptoms,  he  observed  an  erythema  of  the  legs; 
the  chickens  recovered  when  given  whole  corn,  dementi  (711)  fed 
chickens  on  cooked  corn  and  "polenta"  and  observed  a  development 
of  nervous  and  digestive  disturbances,  but  only  after  many  months 
of  feeding.  Szalagyi  and  Kriwuscha  (712)  also  conducted  some  corn 
feeding  experiments  with  chickens,  ducks  and  geese. 

Urbeanu  (713)  carried  out  some  rat  and  chicken  experiments,  and 
noted  some  skin  symptoms  in  both.  In  the  former,  this  was  apparent 
only  after  10  to  12  months  of  feeding,  and  was  favorably  influenced 
by  the  addition  of  potatoes  and  soya  beans.  Sua*rez  (714),  experi- 
menting with  pigeons  and  mice,  observed  in  both  a  disease  similar 
to  beriberi  after  25  to  35  days,  which  was  favorably  affected  by 
yeast.  Experimenting  with  dogs,  Nitzesco  (715)  stated  that  corn 
was  not  a  complete  food.  Rondoni  (716)  attempted  to  use  guinea 
pigs  in  his  pellagra  work,  but  recognized  that  these  animals  were 
unsuited  for  this  purpose,  and  therefore  suggested  that  the  work  be 
continued  on  man  and  monkeys. 

Hogan  (717)  fed  corn  simultaneously  to  rats  and  pigs;  in  the 
former,  this  diet  was  very  nearly  sufficient,  but  in  the  latter,  a  lack 
of  protein  was  noted,  since  on  adding  egg- albumin,  growth  was 
resumed.  This  brings  us  to  a  consideration  of  the  nutritive  value  of 
corn  proteins,  a  subject  which  has  assumed  considerable  significance 
in  the  etiology  of  pellagra.  In  this  connection,  Mendel  and  Fine  (718) 
carried  out  a  series  of  dog  experiments.  According  to  their  results, 
corn  glutellin,  a  preparation  consisting  of  zein  and  glutenin,  seemed 
to  be  as  well  utilized  as  meat.  The  relationship  between  the  two 
protein  components  varied  within  certain  limits,  but  averaged  1:1. 
The  proteins  were  separated  by  means  of  alcohol,  in  which  zein  is 
soluble.  Osborne  and  Mendel  (719)  and  Baglioni  (720)  conducted 
experiments  on  the  nutritive  value  of  corn  protein.  The  utilization 
was  better  with  the  flour  than  with  the  proteins  isolated.  All  three 
products  however  gave  a  nitrogen  retention,  and  it  would  therefore 
be  concluded  that  corn  also  contains  proteins  of  a  higher  nutritive 
value  than  that  of  zein.  McCollum  and  Simmonds  (721)  found  that 
when  rats  are  fed  on  a  mixture  of  80  per  cent  corn  and  20  per  cent 
beans,  there  is  still  evidence  of  a  lack  of  vitamine  A  and  protein. 


252 


THE   VITAMINES 


The  investigations  of  Johns,  Finks  and  Paul  (722)  gave  the  following 
percentage  composition  of  corn  as  regards  various  protein  compounds : 

Globulin 21.9 

Zein 41.4 

Glutellin 30.8 

Alcohol-insoluble  part 5.9 

While  ifc  is  known  that  zein  is  not  a  complete  protein,  the  com- 
position of  glutellin  shows  it  to  be  of  good  nutritive  value.  As  an 
explanation  of  the  poor  nutrition  resulting  from  an  exclusive  corn 
diet,  these  investigators  stated  that  certain  flour  products  lack,  first 
of  all,  vitamine  B;  second,  although  they  contain  complete  proteins, 
they  occur  in  dilute  form,  so  that  the  protein  must  be  added  as  con- 
centrates. It  is  interesting  to  note  that  this  view  corresponds 
exactly  with  our  present  conception  of  the  matter.  Osborne  and 
Mendel  (723)  found  that  the  dietary  deficiency  arising  from  corn 
meal  feeding  is  caused  by  a  deficiency  of  protein.  This  deficiency 
may  be  corrected  by  an  addition  of  skimmed  milk  powder,  which 
must  amount  to  50  per  cent  of  the  corn  fed,  before  the  nutritive 
condition  is  restored  to  normal. 

We  come  now  to  a  consideration  of  the  vitamine  content  of  corn, 
and  how  it  is  affected  by  milling.  It  has  been  known  for  a  long 
time  that  corn  germ- contains  much  more  fat  than  the  whole  corn. 
The  appended  table  is  taken  from  the  work  of  Woods  (724). 
The  protein  content  varies  in  the  same  way. 


FAT 

PKOTEIN 

Whole  corn  

per  cent 

4  3 

per  cent 

12  7 

Endosperm. 

1  5 

12  2 

Husk  

1.6 

6.6 

Germ  

28  6 

21  7 

We  have  personally  made  analyses  of  various  South  Africa  corn 
products  (I.e.  525)  and  concluded  that  milling  causes  considerable 
losses,  which  may  be  prevented.  Our  figures  on  milling  are  recorded 
below. 


VITAMINE    CONTENT   OF   FOODSTUFFS 


Data  pertaining  to  milled  maize   (97  per  cent  of  the  total  grain) 


253 


COLOR  RE- 

NITROGEN 

o 

ACTION. 

PM 

FOLINAND 

• 

rj) 

j 

Q 

MACALLUM 

2 

d 

I 

B 
« 
o 

9 

o 

«• 

« 
• 

B 

o 

PH 

Alcoholic 

M 

| 

a 

•a 

53 

w 

h 

i 

Q 

extract 

B 

l 

a 

02 

b 

a 

H 

! 

9 

"3 
M 

a 

Q 

3 

cj 

M 
PH 

1 

\ 

o 

8 

E 

Hot 

Cold 

1.  Whole 

maize 

grain 

12.71 

1.56 

1.73 

1.74 

0.14 

0.99 

0.56 

4.5 

3.88 

0.247 

0.0154 

0.35 

0.55 

2.  Highly 

milled 

meal  86 

per  cent 

12.63 

1.48 

1.67 

1.73 

0.13 

0.95 

0.36 

3.87 

3.36 

0.22 

0.0098 

0.23 

0.45 

3.  First 

milling 

from  no.  2 

10.48 

2.09 

1.23 

0.14 

0.61 

0.30 

4.66 

4.18 

0.357 

0.0164 

1.00 

1.00 

4.  Second 

milling 

from  no.  2 

10.71 

4.04 

2.31 

2.34 

0.16 

1.40 

1.43 

12.79 

11.09 

0.438 

0.0353 

0.70 

0.60 

5.  Slightly 

milled 

meal,   97 

per  cent 

12.41 

1.60 

1.73 

1.84 

0.12 

1.00 

0.54 

4.21 

3.63 

0.233 

0.0153 

0.30 

0.45 

6.  Bran 

from  5, 

above 

10.55 

1.400.65 

0.07 

0.300.23 

1.86 

1.58 

0.2710.0084 

0.35 

0.70 

We  have  found  nothing  in  the  literature  pertaining  to  the  nutritive 
value  of  the  proteins  of  different  corn  layers.  It  is,  of  course,  possible 
that  the  two  chief  proteins  of  corn — zein  and  glutenin — are  localized 
in  various  layers,  so  that  milling  may  deprive  the  corn  of  glutenin, 
and  hence  enrich  it  with  the  less  valuable  zein.  With  the  above 
analyzed  products,  we  have  carried  out  feeding  experiments  on 
pigeons,  but  have  never  observed  beriberi.  From  the  diagram  "of 
the  corn  kernel  (fig.  54),  it  is  possible  to  see  the  localization  of  the 
germ,  explaining  the  loss  of  substance  and  nutritive  value  during 
milling.  Analyses  of  corn  milling  products  were  also  published  by 
Juritz  (725)  and  by  McCrae  (726).  Poppe  (727)  found,  besides  this, 
that  by  steeping  the  corn,  similar  to  cooking,  the  nutritive  substances 
extracted  by  the  water  amount  to  36.2  per  cent.  McCollum,  Sim- 
monds  and  Pitz  (728)  found  that  corn  contains  all  of  the  necessary 


254 


THE    VITAMINES 


amino  acids,  but  not  in  the  proper  proportion  to  each  other  and, 
in  addition,  was  poor  in  vitamine  A  and  salts.  While  this  was  found 
to  be  the  case  in  rats,  Weill  and  Mouriquand  (729)  were  able  to 
maintain  pigeons  on  whole  corn  for  more  than  240  days,  whereas 
decorticated  or  sterilized  corn  produced  severe  disturbances.  Voegt- 
lin,  Lake  and  Myers  (I.e.  690)  found  that  whole  corn  contains  suffi- 
cient vitamine  B  for  pigeons.  However,  if  they  fed  the  same  corn 
with  the  biggest  part  of  the  aleurone  layer  and  germ  removed  the 
pigeons  and  chickens  developed,  in  about  3  weeks,  a  disease  similar 
to  beriberi  which  could  be  cured  by  giving  vitamine  B.  Voegtlin, 
Sullivan  and  Myers  (730),  as  well  as  Hughes  (731),  also  obtained 
similar  results. 


FIG.  54.  CKOSS-SECTION  OF  CORN  KERNEL 
A,  endosperm.,  B,  husk,  C,  germ 

Steenbock  and  Boutwell  (I.e.  588)  showed,  contrary  to  the  ruling 
view  that  corn  is  poor  in  vitamine  A,  that  certain  strongly  yellow 
colored  varieties  are  quite  rich  in  this  vitamine.  In  connection  with 
this,  we  wish  to  emphasize  once  more  that  corn  is  a  complete  diet  for 
most  species  of  birds.  It  is  different  with  mammals,  for  which  corn 
is  not  so  favorable,  and  this  fact  is  now  generally  attributed  to  the 
low  biological  value  of  the  corn  proteins,  although,  inasmuch  as  corn 
contains  a  protein  that  is  adequate  for  birds,  it  is  incorrect  to  speak 
of  the  low  biological  value  of  corn  proteins.  In  experimenting  with 
mammals,  particularly  those  that  can  develop  scurvy,  we  must  not 
forget  that  corn,  at  least  when  dry,  contains  no  C  vitamine.  This 


V1TAMINE    CONTENT   OF   FOODSTUFFS  255 

minimal  value  of  corn  proteins  does  not  depend  so  much  on  the 
unfavorable  content  of  amino  acids,  as  that  for  some  animals,  the 
dilution  of  the  complete  proteins  by  the  incomplete,  and  by  large 
amounts  of  starch,  has  an  unfavorable  effect  on  the  utilization. 

POTATOES 

We  have  already  stated  that  in  a  population  that  is  nourished 
chiefly  on  potatoes,  no  avitaminoses  appear  and  pellagra  is  unknown. 
The  value  of  potatoes  as  a  protection  against  deficiency  diseases  has, 
to  a  certain  extent,  been  recognized  by  some  investigators.  Although 
the  protein  content  of  potatoes  is  small,  and  although  large  quan- 
tities must  be  used  because  of  the  great  dilution  of  the  more  valuable 
components  present,  it  seems  to  be  certain  that  this  dilution  is  harm- 
less to  people.  The  author  has  often  had  the  opportunity  of  observing 
Polish  peasants  at  their  meals.  When  the  men  returned  from  the 
fields,  an  enormous  plate  of  potatoes  was  set  before  them,  so  that  a 
city  dweller  could  hardly  conceive  of  the  possibility  of  consuming 
such  a  great  amount  of  food.  Still,  the  entire  contents  of  the  dish 
(not  plate),  cooked  with  a  trace  of  lard  to  give  added  taste,  dis- 
appeared in  an  unbelievably  short  time.  From  the  many  modern 
investigations  on  the  nutritive  value  of  potatoes,  it  is  clear  to  us 
why  these  peasants  must  consume  such  an  unusual  amount  of  pota- 
toes. It  is  correlated  with  the  small  content  of  protein,  together 
with  the  three  known  vitamines.  The  splendid  utilization  of  potato 
proteins  was  unquestionably  demonstrated  by  Hindhede  (732),  and 
we  shall  discuss  this  later  on.  His  findings  were  confirmed  by 
Abderhalden,  Fodor  and  Rose  (733)  as  well  as  by  Rose  and  Cooper 
(734).  Despite  the  work  of  Rubner  (735),  we  believe  that  Hind- 
hede's  results  are  on  a  firm  foundation. 

The  experiments  with  animals,  especially  rats  and  guinea  pigs, 
resulted  differently  from  those  on  man.  We  have  already  seen  in 
the  author's  earlier  work  on  guinea  pig  scurvy,  that  fresh  potato 
juice  did  not  possess  any  great  protective  action  against  this  disease. 
McCollum,  Simmonds  and  Parsons  (736),  experimenting  with  rats, 
obtained  results  quite  dissimilar  to  those  in  man.  They  found  that 
potatoes  cannot  take  the  place  of  other  grains  in  their  nutritive  value, 
and  are  deficient  in  salts,  protein  and  vitamine  A.  One  would  be 
tempted  to  conclude  from  this  that,  between  rats  and  man,  there  are 
differences  in  the  utilization  of  potatoes.  Osborne  and  Mendel  (737) 


256  THE    VITAMINES 

found  that  the  vitamine  B  content  of  potatoes  is  sufficient  for  rats, 
and  no  difference  was  apparent  between  new  and  old  potatoes.  Abder- 
halden  and  Schaumann  (I.e.  499)  made  the  observation  that  when 
pigeons  are  fed  on  dried  potatoes,  they  develop  edema  on  the  lower 
part  of  the  legs.  As  regards  the  vitamine  content,  Chick  and 
her  co-workers  (I.e.  333)  found  that  potatoes  (cooked  and  raw) 
contain  all  three  vitamines  in  small  quantities.  Boruttau  (738) 
studied  the  effect  of  temperature  on  the  nutritive  value  of  potatoes 
in  mice.  Similar  experiments  were  made  by  Auer  (739)  in  Hof- 
meister's  laboratory.  The  effect  of  drying  was  studied  by  Givens  and 
McClugage  (I.e.  657). 

MILK 

Milk  is  such  an  important  food  for  life  that  we  must  de- 
scribe it  more  in  detail.  With  regard  to  the  milk  question,  we 
were  first  to  suggest  the  relationship  between  the  composition  of  the 
milk  and  some  deficiency  diseases  in  children.  In  1912,  we  stated 
two  factors,  without  which  the  explanation  of  certain  avitaminoses 
would  be  impossible.  The  two  essentials  are  first,  that  on  heating 
the  milk,  certain  vitamines  become  more  or  less  completely  destroyed, 
and  second,  that  the  vitamine  content  of  milk  must  depend  on  the 
vitamine  content  of  the  fodder.  Although  both  are  self-evident,  they 
gained  recognition  only  with  difficulty.  The  following  rules,  laid 
down  by  us  at  that  time  for  feeding  of  milk  to  children,  are  still  of 
value  at  present :  Hygiene  of  the  cow,  good  fodder,  least  possible  boiling 
of  the  milk,  least  possible  storage  after  milking  and  after  pasteurization. 
Our  uncompromising  attitude  on  the  milk  question  was  made  neces- 
sary by  the  fact  that  Lane-Claypon  (740),  in  a  report  to  the  Local 
Government  Board  in  London,  came  to  the  conclusion  that  boiling 
does  not  affect  the  nutritive  value  of  milk.  As  a  matter  of  fact, 
the  same  investigator  (741),  one  year  later — 1913 — declared  that  the 
question  needed  further  development.  The  ensuing  years  have 
brought  to  light  a  great  number  of  investigations  which  corroborated 
our  view  on  the  subject  completely.  Later,  Lane-Claypon  (742) 
published  a  book  dealing  with  the  question  in  detail. 

Demonstration  of  vitamines  in  milk 

In  1912  we  (I.e.  324),  proved  the  presence  of  vitamine  B  in  milk, 
and  Hopkins  also  discovered  the  presence  of  a  growth  substance. 


VITAMINE    CONTENT   OF   FOODSTUFFS  257 

The  description  of  infantile  beriberi  by  Hirota  (743)  and  Andrews 
(744)  showed,  of  course,  that  the  vdtamines  in  milk  must  originate  in 
the  food  of  the  mother,  since  these  young  ones  were  nursed  by  mothers 
suffering  from  beriberi.  The  question  as  to  the  nutritive  value  of 
heated  milk,  especially  in  relation  to  infantile  scurvy  and  without 
knowledge  of  vitamines,  was  treated  by  Sir  Thomas  Barlow,  Neu- 
mann, Heubner  and  others;  we  shall  discuss  this  work  in  detail  in  the 
chapter  on  scurvy. 

Influence  of  food  on  the  vitamine  content  and  composition  of  milk 

Because  of  the  proof  of  the  variation  in  the  composition  of 
milk,  we  are  in  position  to  explain  some  of  the  divergent  statements 
in  the  literature.  These  facts,  long  known  to  cattle  breeders 
and  milk  producers,  of  course  without  any  knowledge  of  vita- 
mines,  must  be  proven  anew  from  the  viewpoint  of  the  vitamines. 
That  the  milk  may  possess  a  different  nutritive  value  according  to  the 
fodder,  has  been  shown  by  Carr,  Spitzer,  Caldwell  and  Anderson 
(745),  Hart,  Nelson  and  Pitz  (746),  McCollum  and  Simmonds  (747) 
and  Drummond  (748).  A  series  of  interesting  experiments  were 
published  by  Eckles,  Palmer  and  Swett  (749-750) .  The  latter  showed 
that  the  composition  of  milk  is  not  constant;  especially  does  the 
protein  and  fat  content  vary  considerably.  The  recognition  of  the 
variation  in  the  protein  content  of  milk  bears  some  relationship  to  our 
modern  conception  of  pellagra — which  we  shall  discuss  further  under 
the  subject  of  pellagra — since  under  these  conditions  infantile 
pellagra  should  be  more  frequently  encountered,  which  is  not  the 
case.  The  variation  in  the  composition  of  milk  explains  the  results 
of  Chick,  Hume  and  Skelton  (751),  who  found  that  for  the  protection 
of  guinea  pigs  against  scurvy,  a  larger  quantity  of  milk  than  that 
found  by  us  was  necessary.  The  question  as  to  the  origin  of  milk 
vitamines  from  the  food  has  already  been  treated  experimentally. 
McCollum,  Simmonds  and  Pitz  (752)  accepted  this  view  for  the  vita- 
mine  A  and  B  content  of  milk.  Osborne  and  Mendel  (I.e.  326)  ob- 
served no  difference  in  the  vitarnine  B  content  of  summer  and  winter 
milk.  Regarding  vitamine  C,  this  relationship  is  very  apparent. 
Hart,  Steenbock  and  Ellis  (753)  fed  cows  on  dry  and  fresh  fodder. 
While  of  the  milk  resulting  from  the  fresh  fodder,  15  to  50  cc.  daily 
were  necessary  to  protect  guinea  pigs  from  scurvy,  of  that  arising  from 
the  dry  fodder,  50  to  75  cc.  were  necessary  to  produce  the  same  result. 


258  THE    VITAMINES 

These  findings  were  confirmed  by  Hess,  Unger  and  Supplee  (754),  who 
showed  further  that  even  a  3  weeks  feeding  with  dry  and  fresh  food 
was  sufficient  to  show  the  above  variations.  Guinea  pigs  lived  an  aver- 
age of  56  days  on  milk  produced  on  the  dry  fodder,  while  on  the  milk 
resulting  from  the  fresh  fodder  the  animals  lived  more  than  120  days, 
and  after  this  time  showed  only  a  mild  scurvy.  In  the  milk  contain- 
ing more  vitamine  C,  more  calcium  and  phosphorus  and  50  per  cent 
more  citric  acid  were  found.  Dutcher  and  co-workers  (754a)  cor- 
roborated the  above  findings  as  to  the  dependence  of  the  vitamine 
content  of  the  milk  on  the  vitamine  content  of  the  cow's  food. 
According  to  Steenbock,  Boutwell  and  Kent  (I.e.  594)  the  same  is 
likewise  true  of  vitamine  A,  after  only  a  3  weeks'  feeding. 

The  vitamine  content  of  milk 

Inasmuch  as  we  have  seen  that  the  vitamine  content  of  milk  varies 
according  to  the  composition  of  the  diet,5  we  must  add  that  the 
vitamine  content  even  of  raw  milk  is  not  very  high.  Therefore  it  is 
obvious  that  any  treatment  of  milk,  or  merely  storing,  may  easily 
diminish  the  margin  of  safety.  The  author  (I.e.  325)  as  well  as 
Osborne  and  Mendel  (I.e.  326)  and  Gibson  and  Conception  (755) 
showed  that  the  vitamine  content  of  milk  is  not  very  high.  That  this 
is  also  true  of  vitamine  C  was  demonstrated  by  Chick,  Hume  and 
Skelton  (756).  As  regards  vitamine  A,  it  seems  to  occur  in  larger 
quantities;  still,  we  are  rather  uncertain  of  its  stability,  since  every 
manipulation,  because  of  the  apparently  easy  oxidizability  of  the 
substance,  may  have  an  effect  on  the  nutritive  value  of  the  milk. 

We  may  readily  conceive,  from  the  above,  that  the  treatment  of 
milk  may  produce  a  deficiency  disease  in  children  whose  practically 
sole  food  it  is.  That  this  is  the  case  in  infantile  scurvy,  was  shown 
by  Hess  (757)  using  pasteurized  milk.  The  disease  developed  after 
a  few  months  and  was  only  of  a  mild  nature.  Hess  (758)  and  Hess 
and  Unger  (759)  correctly  observed  that,  aside  from  heating,  storage 
plays  a  great  part  in  the  conservation  of  antiscorbutic  power.  Miller 
(760)  was  also  of  the  opinion  that  since  raw  milk  contains  only  a 
small  quantity  of  vitamine  C,  just  within  the  margin  of  safety, 
further  manipulation  may  be  dangerous. 

5  In  addition,  Moore  (754b)  raised  the  question  as  to  whether  infectious 
conditions  of  the  cows  may  not  influence  the  vitamine  content  of  milk. 


VITAMINE    CONTENT   OF   FOODSTUFFS  259 

The  influence  of  heat  on  the  vitamine  content  of  milk 

Practically  all  of  the  studies  related  to  this  point  were  made  with 
vitamine  C.  In  this  respect,  most  workers  agree  that  the  use  of 
heated  milk  may  result  in  scurvy.  Some  investigators,  on  the  con- 
trary, believe  that  the  decrease  in  the  nutritive  value  after  heating 
cannot  be  attributed  entirely  to  the  vitamines.  McCollum  and 
Davis  (I.e.  97)  stated  that  this  is  related  to  an  injury  of  the  casein 
by  heat.  We  have  already  mentioned  this  explanation  and  experi- 
mentally disproven  it.  Daniels  and  Stuessy  (761)  showed  that  rats 
fed  on  milk,  cooked  for  1  minute,  could  not  grow.  This  experiment 
was  repeated,  varying  the  temperature  and  time  of  heating.  They 
found,  at  the  time,  that  an  addition  of  protein,  in  the  form  of  casein 
or  egg-yolk,  permitted  the  rats  to  resume  growth.  In  a  recent  paper, 
Daniels  and  Loughlin  (762)  report  that  the  results  of  Daniels- 
Stuessy  with  casein  and  egg-yolk  were  to  be  attributed  simply  to  an 
addition  of  calcium  salts.  They  found  that  when  milk  is  heated 
slowly  at  a  lower  temperature,  as  in  pasteurization,  a  deposit  con- 
taining calcium  salts  is  formed  in  the  kettle.  If  the  milk  is  quickly 
boiled  up,  or  if  care  is  taken  to  mix  the  above  deposit  in  with  the 
milk,  then  the  rats  grow  just  as  well  as  on  unheated  milk.  The  same 
results  were  obtained  when  soluble  calcium  salts,  like  calcium  glycero- 
phosphate,  were  added  to  the  heated  milk.  Regarding  the  signifi- 
cance of  the  results  of  Daniels  and  Loughlin  critically,  they  maintain 
that  the  decrease  in  the  nutritive  value  of  milk  after  heating  is  due, 
not  to  the  destruction  of  B-  and  A-vitamines,  but  to  the  precipitation 
of  calcium  salts.  Although  the  destruction  of  vitamine  C  is  not 
touched  upon  in  this  work,  the  investigators  wished  to  convey  the 
impression  that  the  changes  of  the  milk  on  cooking  had  nothing  to 
do  with  the  vitamines.  It  is  not  impossible,  however,  that  if  they 
would  repeat  their  work,  using  children,  as  they  proposed  to  do,  the 
results  would  be  different  from  those  with  rats. 

The  first  investigation  on  the  influence  of  heating  on  vitamine  C 
was  made  by  Frolich  (763).  Milk  heated  at  98°  C.  for  10  minutes 
lost  its  protective  action  against  guinea  pig  scurvy,  while  heating  at 
70°  C.  for  30  minutes  gave  uncertain  results.  Whereas  the  author 
found  that  50  cc.  milk  prevented  scurvy  in  guinea  pigs,  Chick,  Hume 
and  Skelton  (I.e.  751)  observed  that  this  was  not  sufficient.  Barnes 
and  Hume  (764)  investigated  the  effect  of  heating,  drying  and  rapid 


260  THE   VITAMINES 

boiling  on  the  vitamine  content  of  milk.  Dried  milk  (dried  rapidly 
at  a  high  temperature)  decreased  markedly  in  its  vitamine  C  content 
and  was  inferior  to  milk  boiled  for  a  short  time.  The  activity  of 
vitamines  A  and  B  seemed  to  be  unaffected  by  drying.  Hart,  Steen- 
bock  and  Smith  (765)  obtained  similar  results  with  raw  and  evapo- 
rated milk. 

Influence  of  drying  and  evaporation  on  milk 

From  an  early  "Report  to  the  Local  Government  Board"  (766), 
we  see  that  vigorous  objections  were  raised  against  the  nutritive 
value  of  dried  milk.  In  particular,  the  dilution  necessary  before 
consumption  was  regarded  as  fraught  with  danger.  This  important 
question  was  officially  investigated  in  the  United  States  and  in 
England,  and  a  report  on  this  subject  appeared  in  the  United  States 
Public  Health  Reports  (767).  Winfield  (768)  studied  this  question, 
using  87  children  and  40  rats,  and  came  to  the  conclusion  that  dried 
milk  seems  to  be  complete,  but  only  for  the  first  growth  period.  The 
rats  did  not  grow  any  more  after  they  reached  two-thirds  of  their 
size,  and  the  usual  mixed  diet  had  to  be  resorted  to  at  this  point. 
Naish  (769)  stated  that  in  the  use  of  dried  skimmed  milk  for  feeding 
children  the  danger  of  scurvy  does  not  exist,  even  without  fruit 
addition,  while  Prit chard  (770)  recommended  the  addition  of  fruit. 
Hess  and  Unger  (I.e.  631)  are  of  the  opinion  that  milk,  dried  according 
to  the  Just-Hatmaker  process  for  a  few  seconds  at  116°C.,  is  protective 
against  scurvy  equally  as  well  as  fresh  milk.  In  a  few  cases,  a  cure 
could  even  be  brought  about.  Sekine  (771)  conducted  some  experi- 
ments with  condensed  milk  and  showed  that  young  mice  remained 
normal  on  this  diet  for  about  100  days,  after  which  they  suffered  from 
beriberi  and  anemia.  On  the  addition  of  iron  and  vitamine  B,  they 
again  became  normal.  Coutts  (772)  found  that  dried  milk  gave 
better  results  in  the  nutrition  of  children  than  did  fresh  milk.  He 
believed  that  3  months  old  infants  could  tolerate  dried  whole  milk. 
He  recommended  also  a  synthetic  milk,  in  which  the  cream  was 
substituted  by  cheaper  vegetable  fats  which  were,  however,  appar- 
ently poor  in  vitamine  A.  Gerstenberger  (773)  and  his  co-workers 
tried  to  replace  the  butter-fat  by  a  cheaper  fat.  They  started  with 
skimmed  milk  to  which  were  added  vegetable  fat  and  cod  liver  oil. 
This  milk  was  fed  to  311  children,  who  subsequently  showed  no  signs 
of  rickets,  spasmophilia  or  anemia. 


VITAMINE    CONTENT   OF   FOODSTUFFS  261 

General  remarks  on  the  nutritive  value  of  milk 

The  question  of  dilution  in  connection  with  the  use  of  condensed 
and  dried  milk  appears  also  to  be  of  some  practical  significance. 
Washburn  and  Jones  (774)  have  shown  that  the  fat  content  of  milk 
is  likewise  of  great  significance.  Working  with  young  pigs,  they 
demonstrated  that  skimmed  milk,  as  such,  does  not  permit  of  growth 
in  these  animals.  Milk  with  a  moderate  amount  of  fat  (2.5  per  cent) 
seemed  to  give  the  best  results,  while  too  much  fat  (5  per  cent) 
produced  rapid  growth,  the  animals  being  too  fat  and  apathetic. 
Too  great  a  dilution  of  the  milk  was  followed  by  a  considerable 
decrease  in  its  nutritive  value,  while  a  very  concentrated  milk  reared 
young  that  were  excessively  fat  and  weak,  the  bones  showing  only 
two-thirds  of  their  normal  strength.  Lately,  views  have  appeared 
which  emphasize  the  exceptional  position  of  milk  in  nutrition. 
Freise  (I.e.  342),  as  a  result  of  some  rat  experiments,  expressed  the 
view  that  the  milk  was  either  lacking  an  important  constituent  or 
had  lost  it  during  manipulation.  He  observed,  however,  that 
animals  on  a  deficient  diet  recovered  rapidly  when  an  addition  of 
10  per  cent  dried  milk  was  given,  so  that  milk  possesses  a  splendid 
supplementary  action.  The  same  characteristic  is  manifested  by 
barley-malt.  Honey,  cereals,  alcoholic  extracts  of  barley-malt, 
carrots,  egg-yolk,  beans,  casein  and  smaller  amounts  of  dried  milk 
(less  than  10  per  cent)  no  longer  show  the  activity  of  dried  milk. 
Mattill  and  Conklin  (775),  working  with  rats,  obtained  the  same 
results.  On  ordinary  milk  the  animals  grew  well  at  first,  but  later, 
especially  in  50  to  100  days  old  females,  growth  ceased.  Autolyzed 
and  ordinary  yeast  and  wheat  germ  were  added,  resulting  in  only 
temporary  improvement;  addition  of  iron  and  casein  was  without 
effect.  On  the  other  hand,  better  results  were  obtained  on  adding 
dried  milk,  showing  that  the  dilution  of  the  foodstuff  given  is  of 
significance.  Dried  milk  was  shown  not  to  be  the  ideal  food  for 
later  stages  of  life.  Here  again,  a  dilution  (55  parts  dried  milk, 
40  parts  starch  and  5  parts  butter)  was  of  help  resulting  in  normal 
growth  and  reproduction.  The  dilution  of  dried  milk  with  lard  gave 
the  same  results  as  with  starch.  Addition  of  1  per  cent  yeast 
resulted  in  normal  growth.  For  these  reasons,  the  authors  raised 
the  question,  whether  the  addition  of  yeast  does  not  introduce  some- 
thing that  is  missing  in  the  other  combinations,  and  proposed  new 


262  THE   VITAMINES 

experiments  to  clear  up  this  point.  From  what  has  been  said  in 
this  chapter,  it  is  evident  that  we  are  not  yet  fully  informed  as  to 
the  nutritive  value  of  milk,  and  that  this  foodstuff  still  presents  some 
riddles  for  us  to  solve. 

MEAT 

The  significance  of  meat  as  a  component  of  our  diet  is  threefold. 
First,  as  an  important  source  of  protein;  second,  as  a  source  of  vitam- 
ines  when  it  is  used  as  a  sole  food;  and  third,  for  its  "vitamine- 
sparing"  action.  The  last  characteristic  will  be  treated  more  fully 
later.  In  accord  with  our  own  experience,  we  know  of  no  single  food 
which  requires  so  little  vitamine  addition  as  does  protein  material, 
especially  meat. 

When  we  speak  of  meat  in  this  chapter,  we  have  in  mind  muscle 
tissue  only,  since  glandular  organs  like  liver,  kidney,  etc.,  are  charac- 
terized by  a  special  richness  in  vitamines.  Vitamine  B  was  first 
demonstrated  in  meat  by  Cooper  (I.e.  535).  Osborne  and  Mendel 
(776)  investigated  the  value  of  dried  meat  and  did  not  find  it  par- 
ticularly rich  in  vitamine;  on  successive  extraction  with  water,  the 
amount  of  vitamine  B  decreased  considerably.  According  to  Osborne 
and  Mendel  (777),  dried  meat  contains  sufficient  vitamine  A,  while 
pig's  brain  and  heart  has  enough  of  both  A-  and  B-vitamines.  Cole 
(778)  reported  that  powdered  meat  contains  a  sufficient  amount  of 
A-  and  B-vitamines. 

Pertaining  to  the  vitamine  C  content  of  meat,  practical  experience 
and  laboratory  findings  do  not  seem  to  go  parallel.  Naturally,  the 
discrepancies  are  only  apparent,  since  practice  has  shown  that  fresh 
meat  contains  small  amounts  of  vitamine  C.  This  is  evident  from 
the  report  of  Nansen  (779)  on  his  polar  expedition,  as  well  as  from  the 
report  of  Stef ansson  (780) .  The  latter  stated  that  scurvy  was  cured 
by  an  addition  of  fresh  meat.  However,  we  already  knew  from  the 
work  of  Curran  (781)  that  2  to  4  pounds  of  raw  meat  are  necessary, 
since  he  observed  cases  of  scurvy  on  a  daily  consumption  of  500  grams 
cooked  meat.  Willcox  (782)  stated  that  in  the  English  army  in 
Mesopotamia  scurvy  developed  because  the  Indian  troops  refused 
to  eat  meat.  He  was  of  the  opinion  that  meat  could  function  as  an 
antiscorbutic.  Pitz  (I.e.  365),  in  his  studies  on  guinea  pig  scurvy, 
found  that  an  increase  in  the  protein  proportion  of  the  diet  definitely 
prolongs  the  life  of  the  animal,  which  may  perhaps  be  regarded  as 


VITAMINE    CONTENT   OF   FOODSTUFFS  263 

the  "vitamine-sparing  action"  of  protein.  Butcher,  Pierson  and 
Biester  (783)  found  that  guinea  pigs  could  not  be  protected  against 
scurvy  by  an  addition  of  fresh  meat  juice.  This  was  also  shown  by 
Givens  and  McClugage  (784)  with  dehydrated  meat ;  however,  these 
results  may  be  criticized  on  the  basis  that  guinea  pigs  need  more 
vitamine  C  than  does  man.  That  the  vitamine  requirements  of 
different  animals  may  vary  has  already  been  demonstrated  by 
Harden  and  Zilva  (I.e.  455). 

Parsons  (I.e.  191a)  showed,  on  the  contrary,  that  some  organs, 
especially  the  liver,  but  also  spleen  and  kidney,  contain  relatively 
large  amounts  of  vitamine  C — so  much  so,  that  10  grams  of  pig's 
liver  per  day  is  sufficient  to  protect  guinea  pigs  from  scurvy.  By 
the  preparation  of  extracts,  instead  of  juices  or  organs  themselves, 
Parsons  was  able  to  add  larger  quantities  of  extractives  from  the 
muscle,  showing  that  an  extract  of  55  to  95  grams  of  muscle  suffices  to 
protect  guinea  pigs  from  scurvy.  Fish  muscle  possessed  no  anti- 
scorbutic property. 

THE   VITAMINE    CONTENT   OF   THE   MOST   FREQUENTLY  USED 
FOODSTUFFS 

The  compilation,  which,  as  we  are  well  aware,  cannot  be  complete, 
gives  the  reader  an  idea  of  the  number  of  investigations  carried  out 
for  the  demonstration  of  the  present  known  facts.  We  have  not 
sought  to  give  the  figures  for  the  vitamine  content  since  we  believe 
that  sufficient  progress  has  not  yet  been  made.  In  the  table,  a  single 
plus  sign  denotes  that  the  respective  vitamine  is  present  only  in 
traces.  With  regard  to  protein,  the  plus  sign  indicates  that  it  is  not 
adequate  for  nutrition.  From  the  table,  we  may  see  that  those  start- 
ing materials  which  are  very  rich  in  vitamines  contain  proteins  of 
high  biological  value. 


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272 


PART  III 

The  Human  Avitaminoses— Conditions  in 
which  the  Vitamines  Play  a  Role 


CHAPTER  I 

BERIBERI 

The  first  stage  of  vitamine  research,  which  we  shall  assign  to  the 
period  between  1910  and  1920,  served  as  the  instructive  period.  The 
vitamine  hypothesis,  which  can  really  no  longer  be  regarded  as  such, 
was  promulgated  during  this  period,  and  will  survive  in  a  more  or 
less  modified  form.  The  stage  of  enthusiasm,  in  which  all  possible 
pathological  conditions  were  regarded  as  deficiency  diseases,  has 
passed  and  we  shall  endeavor  now  to  treat  the  matter  from  the 
objective  point  of  view.  In  the  study  of  the  known  and  also  the 
suspected  avitaminoses,  we  meet  with  the  real  difficulty  that  most 
of  the  medical  men  are  divided  into  two  camps.  One  champions 
the  vit amines,  while  the  other  will  have  none  of  them.  The  resultant 
feud  is  not  very  conducive  to  a  speedy  solution  of  the  questions  that 
interest  us  here,  for,  if  one  has  a  preformed  idea,  then  all  facts  are 
viewed  in  a  light  best  in  accord  with  this  idea.  We  find,  in  addition, 
little  cooperation  between  experimental  investigators  and  clinicians. 
This  would  be  the  more  desirable,  because  most  investigators  have 
very  little  clinical  experience,  while  the  clinician  lacks  the  broad 
scientific  basis  of  the  modes  of  nutrition. 

We  frequently  read  in  clinical  papers,  in  connection  with  the 
description  of  a  possible  avitaminosis,  that  dietary  changes  and 
vitamine  therapy  have  been  resorted  to  without  success.  This 
whole  situation  is  usually  disposed  of  in  a  few  words  and  there  is 
frequently  a  failure  to  report  important  observations.  In  purely 
scientific  investigations,  as  in  the  field  of  physiological  chemistry, 
physiology  or  pharmacology,  it  remains  for  the  author  of  the  par- 
ticular subject  to  prove  his  results.  Very  frequently,  his  facts  inter- 
est us  far  more  than  his  conclusions.  In  the  clinical  work  on  avitam- 
inoses, we  often  see  the  conclusions  of  the  individual  authors  merely 
as  personal  impressions,  without  giving  all  the  particulars,  upon 
which  basis,  perhaps,  a  different  conclusion  might  be  possible.  In  the 
interest  of  progress,  it  would  be  very  desirable  if  in  the  clinical 
investigations  in  this  field,  an  exact  description  of  the  previous  diets, 
as  well  as  of  the  vitamine  therapy,  were  given.  In  this  way,  there 
would  be  the  opportunity,  in  every  individual  instance,  of  forming  a 

275 


276  THE    VITAMINES 

personal  conception  of  the  cases  quoted,  which  is  now  usually  quite 
impossible.  Since  our  first  edition,  no  great  progress  has  been 
made  because  of  the  above  reasons,  in  spite  of  many  publications. 
The  conditions  regarded  at  that  time  as  avitaminoses  are  con- 
sidered as  such  to  the  present  day,  while  the  suspected  avitaminoses 
are  still  as  uncertain  as  before.  The  author,  as  the  originator  of 
this  classification  would  like  to  see  all  these  conditions  demonstrated 
as  avitaminoses;  on  the  other  hand,  it  would  be  worth  while  to  elim- 
inate as  soon  as  possible  all  that  does  not  belong  to  this  complex. 

Even  as  regards  the  definite  avitaminoses,  we  are  well  aware  that 
there  will  always  remain  a  minority  who  will  have  a  different  opinion 
from  ours.  While  we  recorded  all  of  these  divergent  opinions  in  our 
first  edition,  we  believed  it  necessary,  from  the  point  of  view  of  uni- 
formity of  the  subject,  to  avoid  this  at  present.  It  should,  however, 
not  be  thought  that  we  wish  to  ignore  the  opponents  of  the  vitamine 
subject.  This  book  is  intended  to  treat  of  vitamines  and  we  shall 
not  stray  from  its  main  purpose. 

What  has  just  been  said  applies  even  more  to  the  description  of 
those  pathological  conditions  which  cannot  possibly  be  regarded  as 
avitaminoses,  but  in  which  the  presence  or  absence  of  vitamines  may 
play  a  part.  The  consideration  of  these  conditions  in  this  book 
should,  however,  not  be  construed  to  indicate  the  author's  belief 
that  they  may  be  related  to  the  vitamines  directly.  It  is  possible 
that  some  of  the  diseases  to  be  described  here  may  in  the  future  be 
explained  differently.  This  book  merely  pictures  the  possible 
aspects  of  future  vitamine  study,  and  does  not  exclude  another  solu- 
tion of  the  problems. 

The  reader  may  ask  why  an  answer  to  the  above  problems  is  not 
yet  at  hand.  Why  can  we  not  say  whether  or  not  a  certain  condition 
can  be  associated  with  the  vitamines  etiologically?  Apart  from 
the  fact  that  the  vitamines  are  indisputably  essential  for  life,  we 
know  very  little  regarding  their  physiological  significance.  To  be 
sure,  we  see  signs  of  a  generally  diminished  assimilative  ability, 
negative  balances  of  some  important  inorganic  and  organic  constitu- 
ents, and  decreased  resistance  to  infection.  Besides  this,  we  see 
changes  in  most  organs,  in  the  glands  of  internal  and  external  secre- 
tion, and  trophic  changes  in  the  skin.  But  just  how  all  these  con- 
ditions can  be  correlated  with  the  lack  of  vitamines  is  at  present 
unknown  to  us.  The  above  list  of  symptoms  is  so  large  that  we 


BERIBEKI  277 

may  associate  them  with  all  possible  conditions.  If  now  we  con- 
sider that  there  is  the  possibility  that  new  vitamines  may  yet  be 
found,  we  recognize  the  many-sidedness  of  the  possible  vitamine 
aspects. 

The  investigation  of  the  avitaminoses  is  rendered  more  difficult, 
in  that  the  symptoms  characteristic  of  these  conditions  may  be 
attributed  to  entirely  different  causes,  aside  from  the  deficiency  of 
vitamines.  Thus,  we  see  the  development  of  edema,  resembling  very 
much  the  hunger  edema,  which  is,  however,  not  of  dietetic  but  of 
infectious  origin.  We  note  beriberi-like  symptoms,  not  related  to 
lack  of  vitamine  B  but  to  toxins  and  poisons;  this  is  also  true  of 
many  trophic  appearances.  Because  of  these  complications,  the 
literature  on  deficiency  diseases  has  become  very  clouded,  since  it 
frequently  happens  that  the  finding  of  a  case  explainable  by  some- 
thing other  than  vitamine  deficiency  is  often  followed  by  the  attempt 
to  discredit  the  vitamines  entirely.  For  these  reasons,  we  must  be 
careful  in  judging  the  etiology  of  sporadic  and  isolated  cases;  it  may 
easily  be  associated  with  an  error  in  diagnosis.  If  such  cases  occur 
in  great  number  (as  is  usually  found  in  practice)  then  a  mistaken 
diagnosis  can  scarcely  be  made. 

The  vitamines  have  lately  been  associated  with  orthopedic  surgery, 
certain  .eye  troubles,  such  as  hemeralopia  and  ophthalmia;  some 
infections  such  as  tuberculosis,  pneumonia,  leprosy;  some  metabolic 
diseases  such  as  arthritis,  diabetes  (also  cancer) ;  and  with  dental 
caries.  We  know  now  that  these  conditions  have  some  relation  to 
the  diet  but  whether  they  are  specifically  associated  with  vitamines 
has  not  yet  been  shown,  and  is  in  fact  difficult  to  do  so.  Since  many 
of  these  pathological  conditions,  in  spite  of  tiresome  research,  have 
given  no  real  explanation,  we  see  no  harm  if  vitamine  research  will 
give  a  new  impulse  to  the  study  of  these  questions.  We  see  many 
examples  of  how  a  misleading  hypothesis  has  led  to  a  favorable 
solution  of  some  questions;  at  any  rate,  if  a  subject  remains  quiescent 
it  can  never  lead  to  progress. 

In  the  description  of  beriberi,  we  shall  dwell  exclusively  on 
the  human  type,  since  many  phases  of  this  question  have  already 
been  treated  in  avian  beriberi.  Above  all,  we  recommend  to  the 
reader  who  is  particularly  interested  in  beriberi  the  fundamental 
work  of  Vedder  (838)  as  well  as  to  contributions  by  Castellani  and 
Chalmers  (839)  and  Schilling  (840),  who  treat  the  subject  from  the 


278  THE   VITAMINES 

viewpoint  of  the  modern  conception  of  beriberi .  Beriberi  is  a  disease 
which,  according  to  Findlay  (841),  has  been  known  for  hundreds,  it 
not  for  thousands,  of  years.  Its  occurrence  was  described  in  the 
Roman  army  which  invaded  Arabia  in  24  B.C.  Chinese  writers  in 
the  second  century  referred  to  this  point.  Neiching,  the  oldest 
medical  book  (2697  B.C.),  speaks  definitely  of  this  disease. 

East  Asia,  including  the  group  of  Polynesian  Islands,  is  the  main 
beriberi  zone.  The  second  greatest  area  is  in  Brazil  and  the  neigh- 
boring countries.  Furthermore,  the  African  coast  shows  numerous 
foci.  It  was  commonly  held  that  a  moist  warm  climate  was  especially 
conducive  for  the  development  of  the  disease  This  report  is  incorrect 
as,  for  example,  in  the  Russo-Japanese  War,  a  severe  outbreak  of 
beriberi  was  noted  in  the  Japanese  army  in  spite  of  the  severe  winter; 
beriberi  is  endemic  also  in  northern  Japan.  The  above  mentioned 
statement  is  based  largely  upon  the  fact  that  rice  grows  well  only  in 
moist  heat.  Japan  is  the  most  afflicted  country.  Balz  and  Miura 
(842)  estimated  the  number  of  beriberi  patients  at  about  50,000 
yearly.  China  is  very  much  less  afflicted,  though  in  Korea  we  find 
numerous  foci.  Furthermore,  we  find  the  disease  quite  well  spread 
over  the  Malay  Peninsula.  The  number  of  cases  there  was  estimated 
by  Fraser  (843)  in  1911  to  be  5540,  among  which  695  deaths  occurred. 
This  is  true  also  of  the  islands  of  the  Malay  Archipelago,  Java, 
Borneo,  Celebes,  the  Molucca  Islands,  New  Guinea,  and  especially 
Sumatra.  Simpson  (844)  reported  cases  in  Singapore,  where  the 
disease  was  often  noted  among  Chinese  coolies.  In  Hawaii,  New 
Caledonia  and  North  Australia,  the  disease  is  mostly  seen  in  Japanese 
and  Chinese  immigrants.  More  beriberi  zones  are  found  in  Cochin- 
China,  Siam  (according  to  Hepburn,  845)  and  Burma,  at  the  mouth 
of  the  Ganges  (according  to  Mulvany,  846),  on  the  east  coast  of  the 
East  Indies,  Ceylon,  Malabar,  and  less  so  on  the  Koromandel  Coast. 

In  Africa,  numerous  foci  exist  on  the  east  and  west  coasts.  Thus, 
Chevalier  (847)  reported  on  an  outbreak  in  Serenli  (East  Africa)  with 
112  cases  in  six  months,  with  44  deaths.  In  the  Congo,  the  rice 
eaters  are  often  attacked  by  the  disease,  while  other  natives,  to  whom 
rice  feeding  is  unknown,  remain  free  from  the  disease.  Dubois  and 
Corin  (848)  describe  a  number  of  cases  in  Bokala  (Belgian  Congo) 
which  were  caused  by  the  consumption  of  "maniok."  Similarly, 
beriberi  is  known  in  Madagascar,  and  in  the  Mauritius  and  Reunion 
Islands. 


BERIBERI  279 

The  Philippines  and  Brazil  form  two  important  foci.  From  Brazil, 
there  is  a  report  of  Lovelace  (849),  who  observed  934  cases  within 
four  years;  the  disease  is  known  there  for  250  years.  The  beriberi 
zone  broadens  out  to  the  north,  till  Rio  de  la  Plata.  Lovelace  is 
not  the  only  observer  in  Brazil;  we  have  also  the  reports  of  Wolcott 
(850)  and  Fraga  (851),  the  latter  having  seen  cases  in  Bahia.  Riddel, 
Smith  and  Igravidez  (852)  described  cases  in  the  military  hospital 
at  Porto  Rico.  From  the  previous  chapter,  we  saw  that  it  did  not 
require  a  consumption  of  white  rice  to  produce  the  disease.  Little 
(853)  saw  a  number  of  cases  in  Newfoundland  resulting  from  the 
ingestion  of  white  flour.  In  the  United  States,  cases  of  beriberi  are 
very  rare  occurrences.  Parker  (854)  describes  an  outbreak  of  this 
disease  in  the  prison  at  Elizabeth,  N.  J.,  as  a  result  of  eating  white 
bread.  Reed  (855)  saw  cases  in  California  among  Chinese  and 
Japanese.  Travis  (856)  described  cases  in  Eddyville,  Kentucky. 

In  Europe,  beriberi  was  observed  in  lunatic  asylums  in  Ireland  in 
1894,  1896  and  1897;  in  1894,  there  were  147  cases  in  Richmond 
Asylum  near  Dublin,  with  25  deaths.  Chantemesse  and  Ramond 
(857)  reported  an  outbreak  of  beriberi  in  the  lunatic  asylum  in  Saint- 
Lemmes  (Angers),  in  1898,  resulting  in  40  deaths.  Schiiffner  (858) 
believed,  in  general,  that  beriberi  cases  occurred  sporadically  from 
time  to  time  and  were  treated  under  another  name.  Recently, 
cases  were  observed  by  Martinez  (859)  in  Spain  and  Portugal.  We 
recognize  from  this  survey  that  beriberi  occurs  in  places,  such  as 
prisons,  hospitals,  etc.,  where  the  food  is  prepared  in  quantities  in 
large  kettles.  A  further  opportunity  for  the  occurrence  of  beriberi 
is  provided  by  wars.  In  the  Russo-Japanese  war,  the  Japanese, 
according  to  Schilling  (I.e.  840),  had  about  70,000  to  80,000  cases, 
while  the  Russians,  as  we  shall  see  later,  had  only  scurvy  cases.  In 
the  last  war,  we  have  similar  observations.  Sicard,  Roger  and 
Rimbaud  (860),  Roger  (861)  and  Leggate  (862)  reported  on  numerous 
cases  among  Chinese  and  Indo-Chinese  laborers,  who  were  trans- 
ported to  France  for  labor  during  the  war.  Massalongo  (863) 
described  cases  on  the  Italian  front.  Willcox  (864),  from  the 
Mediterranean  area  in  the  English  army  in  1916,  and  later  also  from 
Mesopotamia  (865),  reported  cases  which  could  not  be  attributed  to 
rice  consumption.  Braddon  (866)  investigated  a  number  of  cases  in 
the  Mediterranean  war  zone.  Hehir  (867)  and  Sprawson  (868) 
observed  cases  during  the  siege  of  Kut-el-Amara. 


280 


THE   VITAMINES 


MODE    OF   OCCURRENCE    OF   AND   DIETS   LEADING   TO   BERIBERI 

It  is  very  important  for  the  proper  understanding  of  the  nature  of 
beriberi  to  know  the  composition  of  the  diet  leading  to  its  occurrence. 
This  is  the  more  important  since  a  number  of  investigators  have 
reached  the  conclusion  that  the  diet  of  their  patients  was  completely 
satisfactory,  from  the  viewpoint  of  nutritional  studies.  As  for  this, 
we  may  bring  to  our  support  the  slight  amount  of  exact  data  available 
from  two  sources.  On  the  one  hand,  we  know  the  composition  of  the 
diet  in  a  few  instances  where  the  disease  has  been  produced  experi- 
mentally in  man;  on  the  other,  we  have  the  reports  of  a  few  beriberi 
outbreaks,  in  which  the  composition  of  the  diet  was  accurately  noted. 

In  the  first  series,  there  is  the  work  of  Fraser  and  Stanton  (I.e.  47), 
who  produced  beriberi  in  healthy  Japanese  coolies  on  a  diet  made  up 
almost  exclusively  of  white  rice.  Here,  80  to  90  days  passed  before 
the  disease  made  its  appearance.  Another,  and  far  more  important, 
investigation  was  that  of  Strong  and  Crowell  (869).  They  conducted 
experiments  with  volunteers  from  a  prison  in  the  Philippines.  The 
subjects  were  divided  into  three  groups,  each  group  receiving  a 
certain  amount  of  fish,  lard,  bananas,  potatoes  and  sugar.  Rice,  in 
three  different  forms,  was  the  chief  component  of  the  diet. 


NUMBER  OF 

NUMBER  OF 

VOLUNTEERS 

BERIBERI  CASES 

1 

White  rice  and  polishings 

8 

2 

2 

White  rice 

17 

13 

3 

Unpolished  rice 

7 

1  (mild  form) 

From  this,  it  is  evident  that  unpolished  rice  can  practically  prevent 
the  occurrence  of  beriberi,  though  the  vitamine  B  content  of  this 
kind  of  rice  barely  exceeds  the  necessary  amount.  The  first  symp- 
toms were  noted  after  60  days.  Caspari  and  Moszkowski  (870) 
reported  an  experiment  upon  Moszkowski  himself.  He  lived  230 
days  on  white  rice  and  other  vegetable  food,  without  meat,  eggs  or 
cheese,  and  developed  mild  beriberi,  but  with  cardiac  symptoms  and 
considerable  nitrogen  loss.  On  the  addition  of  rice  polishings,  the 
symptoms  promptly  disappeared.  It  seems  likely  that  that  diet 
contained  small  amounts  of  vitamine  B,  since  the  development  of  the 
disease  was  so  long  delayed.  Fraga  (871)  conducted  an  experiment 
with  nine  prisoners.  He  was  unable  to  produce  the  disease  with 


BERIBERI  281 

sterilized  rice  and  sterilized  beans  during  a  period  of  43  days;  the  sub- 
jects objected  to  the  food,  and  the  experiment  had  to  be  terminated. 

The  second  group  of  the  accessible  data  pertains  to  the  composition 
of  the  diet  leading  to  beriberi.  Chamberlain  (872)  investigated  the 
personnel  of  the  native  Philippine  Scouts  comprising  5200  men.  Of 
these,  618  developed  beriberi  in  1908,  and  558  in  1909.  The  food 
leading  to  this  outbreak  consisted  of  340  grams  fresh  beef  or  its 
equivalent  of  bacon,  canned  meat  or  fish,  560  grams  white  rice,  225 
grams  flour  or  bread  and  225  grams  potatoes  or  onions.  When  the 
diet  was  improved  in  1910  by  the  addition  of  more  beans  and  unpol- 
ished rice,  the  number  of  cases  decreased  to  6.  The  cases  described 
by  Riddel,  Smith  and  Igravidez  (I.e.  852),  in  Porto  Rico,  developed 
after  three  months  on  a  diet  consisting  of  rice,  vegetables  and  canned 
meat.  From  the  report  of  Hehir  (I.e.  867),  we  see  that  there  were 
numerous  cases  of  beriberi  among  the  English  troops  at  Kut-el-Amara 
on  a  diet  of  horse-meat,  canned  meat  and  white  flour.  The  siege 
lasted  over  4J  months,  and  this  time  sufficed  for  the  development  of 
the  disease. 

In  confirmation  of  the  author's  findings  (I.e.  559)  and  those  of 
Braddon  and  Cooper  (I.e.  561)  on  pigeons,  Braddon  (I.e.  866)  made 
some  observations  in  the  Mediterranean  war  zone  on  the  dependence 
of  beriberi  on  the  composition  of  the  food.  This  investigator  drew 
up  a  table,  which  we  shall  reproduce,  giving  the  composition  of  two 
diets  capable  of  producing  beriberi,  compared  with  two  other  diets 
which  did  not  result  in  beriberi.  The  content  of  vitamine  B  was 

V 

calculated  on  the  basis  of  100  for  wheat  germ.     The  quotient  ^ 

2L 

shows  the  relationship  between  foodstuffs  containing  and  those  lack- 
ing vitamine  B. 

This  method  of  calculation  is  obviously  not  very  exact,  but  it 
gives  us  an  insight  into  the  relationship  between  protein  and  carbo- 
hydrates; in  beriberi-producing  diets  the  quotient  was  much  smaller. 
It  is  also  interesting  to  note  that  Braddon,  although  the  individuals 
were  still  in  good  condition,  suspected  beriberi,  because  of  the  lack 
of  reflexes,  a  suspicion  which  was  later  confirmed  This  showed 
definitely  that  beriberi  may  be  recognized  prior  to  the  appearance  of 
characteristic  symptoms. 


282 


THE    VITAMINES 


FOODSTUFF 

VITAMINE 
VALUE 

BERIBERI- 
PRODUCING  DIETS 

SATISFACTORY  DIET& 

*Diet  Ai 

Diet  A2 

DietB 

DietC 

Rice  

800 
6400 

400 
400 

400 
8400 

6400 

400 
400 

400 

7600 

600 
3200 
200 
200 
400 
400 
575 
200 

5750 

800 

4800 
200 

100 

5875 

White  bread 

Jam  

Sugar  

Cheese  . 

Dried  fruit 

Salt  fish  

Margarine,  butter  or  oil  

X  =  Total  quantity  of  food  lack- 
ing in  vitamine  B 

Oatmeal  

10 
10 
50 
5 

1200 
400 
1600 

1200 
800 
400 

2400 

400 
850 
400 
1200 

2850 

400 
1200 
600 
800 

3000 

Fresh  meat  or  bacon  

Peas,  beans  or  lintels       .    . 

Potatoes  or  fresh  vegetables  

V  =  Total  wgt.  of  foodstuffs  con- 
taining vitamine  B  

V 

Ratio  —   —  . 

0.2 

0.3 

0.5 

0.5 

*  Weekly  amount  of  food. 


THE  SYMPTOMATOLOGY  OF  BERIBERI 

There  is  no  doubt  that  a  latent  form  of  beriberi  does  exist  (Braddon, 
I.e.  866).  Beriberi  symptoms  are  not  infrequently  noted,  acute 
for  the  most  part,  after  a  trauma  or  over-exertion.  After  an  opera- 
tion, severe  fatal  manifestations  occur  in  persons  who,  before  the 
operation,  showed  no  symptoms  at  all.  Acute  cases  of  beriberi  occur 
in  soldiers  after  a  long  march;  light  abortive  cases  are  very  common. 
The  patients  complain  of  a  feeling  of  heaviness  in  the  legs  and  of 
unsteadiness  in  the  knees  on  walking  or  standing,  so  that  they 
frequently  collapse.  This  occurs  chiefly  in  summer  months.  Very 
often  they  complain  of  palpitation  of  the  heart  after  an  exertion,  and 
of  tingling  and  numbness  in  the  skin  of  the  legs.  Skin  symptoms  in 
the  form  of  erythema,  petechiae,  and  exanthema  are  noted.  Hem- 
eralopia  belongs  to  the  earlier  symptoms;  the  old  therapy  (liver)  of 
Hippocrates  attains  thereby  a  scientific  explanation.  Hemeralopia 
belongs  also  to  the  symptomatology  of  scurvy  and  pellagra,  and  may 


BERIBERI  283 

be  thought  of  as  a  symptom  of  vitamine  hunger.  A  decrease  of  sexual 
ability  is  frequently  observed. 

Hepburn  (I.e.  845)  enumerates,  in  observations  of  100  cases,  the 
following  symptoms:  The  first  symptoms,  as  a  rule,  consisted  in 
one  of  three  manifestations,  (1)  edema  of  the  feet  and  legs  in  50  per 
cent  of  all  cases;  (2)  numbness,  irritability  and  tingling  of  the  legs 
in  42  per  cent,  and  (3)  epigastric  manifestations  in  7  per  cent.  A  soft 
mitral  systolic  murmur  was  noted  in  20  cases  in  the  first  investigation. 
Cardiac  arrhythmia  was  noted  in  5  cases  in  the  first  stage.  Kato 
and  Yamada  (873)  noted  arrhythmia  in  the  convalescent  stage.  The 
body  temperature  was  found  subnormal  by  Doyle  (874) .  As  regards 
the  blood  picture,  Chun  (875)  found  an  increase  in  the  number  of 
leucocytes  amounting  to  100  per  cent.  The  blood  pressure  was 
found  to  range  up  to  200  mm.  by  Yoshikawa,  Yano  and  Nemoto 
(876).  Jida  (877)  investigated  the  cerebrospinal  fluid.  In  a  number 
of  cases  he  found  a  large  pressure,  particularly  in  the  acute  stage, 
up  to  280  mm.  with  a  low  blood  pressure.  This  fluid,  in  cases  with 
high  pressure,  injected  into  the  ear  of  a  rabbit  showed  a  powerful 
vaso-constrictor  action  (which  indicates,  perhaps,  a  high  adrenaline 
content).  Sicard  and  Roger  (878),  on  the  contrary,  failed  to  dis- 
cover any  peculiarities  in  the  cerebrospinal  fluid. 

The  picture  of  beriberi  shows  many  different  forms,  according  to 
Balz  and  Miura. 

1.  The  light,  sensory-motor  form. 

2.  The  dry,  atrophic  form. 

3.  The  wet,  atrophic  form. 

4.  The  pernicious,  acute  form. 

These  forms  may  also  merge  into  one  another.  As  a  rule,  the 
picture  of  the  disease  shows  paralysis  and  muscle  atrophy  in  chronic 
cases;  in  acute  cases,  however,  there  are  gastro-intestinal  disturb- 
ances, anasarca,  hydropericardium  and  severe  cardiac  symptoms. 

1.  The  sensory-motor  form 

This  is  the  most  common  form  of  beriberi.  After  an  exertion, 
especially  during  the  summer  months,  the  patient  feels  unsteady  and 
weak  in  the  legs;  often  there  is  numbness  of  the  arches  of  the  feet 
and  of  the  legs;  the  calf  muscles  are  sensitive  to  pressure.  In  addi- 
tion, there  is  palpitation  of  the  heart  after  exertion,  and  edema  of  the 


284 


THE   VITAMINES 


FIG.  55.  ATROPHIC  BERIBERI  (BALZ-MIURA) 


BERIBERI 


285 


FIG.  56.  SAME  AS  FIG.  55,  BACK  VIEW  (BALZ-MIURA) 


286 


THE    VITAMINES 


BERIBERI  287 

legs.  The  knee  reflex  is  increased  at  first,  and  later  usually  entirely 
absent;  the  temperature  remains  normal.  In  this  stage  a  cure  may 
result  but  it  may  go  over  into  the  dry  atrophic  or  the  wet  atrophic 
form. 

2.  The  dry  atrophic  form  (figs.  55-56) 

In  this  stage,  there  is  general  paralysis  and  muscle  atrophy;  at 
first  the  legs,  then  the  thighs,  hands,  arms,  and  finally  the  back  muscles 
are  involved.  The  patients  then  show  a  skeleton-like  emaciation. 
In  this  stage,  too,  improvement  may  take  place  under  certain  con- 
ditions, and  in  the  end,  after  many  months,  a  complete  cure  may 
result.  The  motility,  electrical  reaction,  the  original  musculature, 
the  patellar  reflex — all  gradually  return.  In  other  cases,  reoccur- 
rences appear,  the  paralysis  and  atrophy  increase,  the  foot  remains 
in  the  position  of  pes  equinovarus  (Fig.  57) ;  the  hands  hang  weakly 
with  half -closed  fingers,  as  in  any  radial  paralysis.  After  long 
prevalence  of  the  disease,  permanent  contractions  of  the  foot  de- 
velop, less  frequently  of  the  fingers  and  biceps.  The  patient  can 
only  walk  on  tip-toes  with  the  help  of  crutches.  These  severe  chronic 
conditions  are  incurable. 

3.  The  wet  atrophic  Jorm 

In  this  condition,  besides  the  above  symptoms,  the  disturbance  of 
circulation  is  more  sharply  prominent.  There  is  palpitation  of  the 
heart,  tachycardia,  dyspnea,  oliguria,  pain  in  the  calf  muscle  and 
edema.  Over  the  ankle,  there  is  a  pale  edema  and  in  severe  forms 
also  in  the  thighs  and  in  the  back,  especially  in  the  shoulder  and 
neck  region;  often  also  in  the  arms  and  on  the  face.  Hydropic 
condition  of  the  serous  cavities,  namely,  the  pericardial  and  the 
pleural,  belongs  to  the  most  important  symptoms.  The  elimina- 
tion of  urine  is  greatly  decreased,  often  to  200  to  300  cc.  or  even  less. 
The  urine  has  a  high  specific  gravity,  is  protein  free,  urates  are 
precipitated  out,  and  it  shows  a  marked  indican  reaction.  If  a  cure 
is  initiated,  the  edema  disappears  with  marked  diuresis,  whereupon 
the  great  emaciation  and  muscle  atrophy  become  apparent.  In  nu- 
merous cases,  death  follows,  partly  because  of  the  severe  hydroperi- 
cardium,  partly  through  paralysis  of  the  heart  muscle,  or,  finally, 
through  a  merging  into  the  acute  pernicious  form.  In  puerperal 
beriberi,  the  wet  form  is  noted,  as  a  rule.  In  severe  cases,  there  is 


288 


THE    VITAMINES 


also  paralysis  of  the  intercostal  muscle,  the  diaphragm,  and  less 
frequently  of  the  cephalic  nerves,  optic,  abducens,  facial  and  hypo- 


FIG.  58.  PARALYTIC  FORM  OF  BERIBERI  (FRASER) 


glossal    nerves.     Usually,    the    laryngeal    muscles    are    paralyzed. 
Paralysis  of  the  vagus  is  generally  regarded  as  the  cause  of  death. 

Epidemic  dropsy.     Under  this  name,  Greig  (879)  described  a  disease 
which  appeared  to  result  from  rice  feeding,  and  which  was  regarded 


BERIBERI  289 

at  first  as  identical  with  wet  beriberi.  It  is  likely  now,  however,  that 
epidemic  dropsy,  sometimes  accompanied  by  fever,  is  related  not  to 
beriberi  but  to  hunger  edema.  Greig  (880)  later  investigated  the 
mode  of  development  of  the  disease  somewhat  more  closely,  and 
found  that  some  natives  feed  on  wheat  from  which  the  husk  and  bran 
are  carefully  removed.  Addition  of  meat  in  these  cases  exercised  a 
beneficial  influence.  The  question  of  the  etiology  of  this  disease  will 
be  taken  up  again  later. 

4.  The  acute  pernicious  or  cardio-vascular  form 

This  variety  can  occur  either  primarily  or  secondarily  from  the 
above  described  forms  by  over-exertion,  after  an  operation,  and  also 
without  apparent  causes.  Quite  suddenly,  often  in  but  a  few  hours, 
a  severe  picture  develops:  precordial  pains,  pain  in  the  epigastrium, 
dyspnea,  tachycardia,  nausea,  vomiting,  diarrhea  and  staggering. 
The  temperature  remains  normal  and  the  mind  clear.  The  heart  is 
markedly  enlarged,  especially  the  right  ventricle;  the  whole  heart 
region  pulsates,  due  to  paresis  of  the  intercostal  muscle  and  the 
increased  heart  labor.  The  visible  pulsation  in  the  epigastrium 
belongs  to  the  early  symptoms  of  the  disease.  Frequently,  there 
develops  a  systolic  murmur  at  the  apex,  and  also  in  the  second  or 
third  intercostal  space;  the  second  heart  sound  is  strengthened.  In 
the  arteria  cruralis,  we  hear  an  arterial-diastolic  murmur;  in  severe 
cases,  even  a  tone.  The  pulse  is  accelerated,  120;  in  severe  cases, 
130  and  140,  full  and  soft.  The  breathing  is  wild  and  wheezing, 
the  whole  thorax  is  raised  spasmodically.  Hoarseness  or  aphonia 
frequently  develops.  The  quantity  of  urine  is  greatly  diminished, 
100  to  200  cc.  per  day,  and  even  less.  The  urine  gives  a  marked 
indican  reaction;  in  some  cases  it  contains  some  albumin,  and  occa- 
sionally it  gives  a  diazo  reaction.  Death  occurs  with  a  small  frequent 
pulse,  cyanosis  and  edema  of  the  lungs,  sometimes  in  but  a  few  days 
or  weeks,  but  sometimes  very  suddenly  in  a  few  hours. 

The  mortality  in  beriberi  varies  within  wide  limits.  In  some  out- 
breaks in  Sumatra,  Java  and  Manila,  the  death  rate  was  frightful — 
up  to  60  to  70  per  cent  and  even  higher,  while  among  the  Dutch 
troops  in  Insulinda  2  to  6  per  cent,  and  among  the  Japanese  troops 
2  to  4  per  cent.  The  death  rate  in  the  wet  form  is  greater  than  in 
the  dry,  and  highest  in  the  acute  pernicious  form.  Formerly,  the 
average  death  rate  was  calculated  to  be  37  per  cent,  but  in  recent 


290 


THE    VIT AMINES 


years,  following  a  better  understanding  of  the  causes  and  the  therapy 
of  the  disease,  there  has  been  a  marked  decrease  in  the  rate.  A 
portion  of  the  chronic  cases  die  with  complications,  chiefly  tuberculosis. 


FIG.  59.  HYDROPIC  FORM  OF  BERIBERI  (FRASER) 

Beriberi  in  infants 

This  form  was  first  described  by  Hirota  (I.e.  743),  and  develops  in 
infants  nursed  by  mothers  suffering  from  beriberi ;  usually,  the  latter 
exhibit  no  signs  of  the  disease.  The  observed  symptoms  are  similar 
to  those  of  adults:  persistent  chronic  vomiting,  cyanosis  around  the 


BERIBERI  291 

mouth  and  nose,  dyspnea,  tachycardia,  aphonia,  edema  of  the  face 
and  the  legs,  oliguria,  but  no  fever.  In  some  of  the  cases,  the  mother 
is  still  in  the  latent  stage  and  shows  only  indicanuria,  while  more 
marked  symptoms  develop  later.  These  statements  were  confirmed 
by  Andrews  (I.e.  744)  in  the  Philippines.  He  found  that  a  great 
number  of  infants  died  there  because  of  this  reason.  The  milk  of 
such  mothers  was  shown  to  be  poor  in  fat  (2  per  cent  instead  of  4  per 
cent),  while  the  calcium  and  phosphorus  content  was  increased.  If 
this  milk  was  given  to  young  dogs,  they  developed  beriberi.  The 
question  as  to  the  mode  of  development  and  the  nature  of  infantile 
beriberi  was  also  treated  by  Lagane  (881),  Weill  and  Mouriquand 
(882)  and  Walshe  (883).  In  addition,  Ogata  and  Jizuki  (884) 
investigated  the  influence  of  beriberi  in  the  mother  on  the  fetus. 
According  to  Roxas  (885),  the  disease  in  the  Philippines  occurs  less 
frequently  in  artificially  fed  children.  The  great  number  of  cases  of 
infantile  beriberi,  compared  with  infrequency  of  scurvy  and  pellagra 
in  naturally  nourished  infants,  makes  it  evident  that  the  formation 
of  vitamine  B  in  the  milk  proceeds  somewhat  differently  from  that 
of  the  other  vitamines. 

PATHOLOGY   OF   BERIBERI 

General 

Diirck  (886)  has  written  a  good  monograph  on  the  subject.  For 
the  most  part,  sections  were  made  of  acute  cases.  The  skin  of  the 
beriberi  corpse  is  quite  edematous,  cyanotic  at  the  extremities,  lips 
and  ears,  often  with  subcutaneous  hemorrhages;  the  musculature  is 
also  edematous.  One  of  the  most  common  typical  findings  is  hydro- 
pericardium,  which  develops  as  a  result  of  the  degeneration  of  the 
nerves  of  the  pericardium.  Hydrothorax,  ascites  fluid  and  pin- 
point hemorrhages  are  frequently  observed  in  the  serous  membranes. 
In  acute  cases,  ecchymoses  are  not  infrequently  noted  in  the  stomach 
and  in  the  duodenum.  The  heart,  especially  the  right  ventricle  and 
auricle,  is  markedly  dilated  and  hypertrophied.  The  heart  muscle 
itself  shows  yellowish  spots  and  streaks  (of  a  fatty  nature).  The 
lungs  are  usually  edematous.  The  kidneys  are  full  of  blood,  but  here, 
as  in  other  organs,  there  are  small  hemorrhages.  Nephritis  was  not 
observed. 


292  THE    VITAMINES 

The  most  important  changes  in  beriberi  were  noted  in  the  periph- 
eral nerves  and  muscles.  In  the  peripheral  nerves,  no  macroscopic 
changes  were  apparent,  aside  from  the  occasional  observation  of 
minute  hemorrhages  in  the  sheath  or  between  the  fibers.  Micro- 
scopically, however,  all  stages  of  degeneration  are  noted.  The 
changes  begin  with  the  nerve;  the  medulla  becomes  varicose,  espe- 
cially near  the  constrictions;  it  falls  apart  in  clumps  and  masses  and 
finally  in  a  granular  detritus.  The  axis  cylinder,  on  the  contrary, 
frequently  remains  intact  for  a  long  time,  but  in  the  end  this  too  dis- 
integrates. The  Schwann's  sheath  then  becomes  a  shapeless  mass, 
with  increase  of  the  nuclei.  In  fresh  acute  cases,  the  number  of 
degenerated  fibers  is  very  slight.  At  the  nerve  endings  in  the 
muscles,  only  those  nerve  fibers  leading  to  the  degenerated  muscle 
fibers  are  swollen  and  granulated.  This  degenerative  neuritis  occurs 
chiefly  in  the  ulnar,  radial  and  medial  nerves  in  the  legs  and  later  in 
the  phrenic  and  vagus  nerves.  Balz  (887)  found  the  same  to  be  true 
of  the  kidney  nerves.  Most  apparent,  however,  was  the  degeneration 
of  almost  all  of  the  spinal  cord  nerves.  Balz  (I.e.  887),  as  well  as 
Ellis  (888),  found  changes  in  the  sympathetic  and  especially  in  the 
cardiac,  pulmonary,  splanchnic  and  solar  plexi.  Rumpf  and  Luce, 
(889)  found  degenerative  processes  in  the  dorsal  roots.  In  the  brain 
and  spinal  cord,  as  well  as  in  other  organs,  there  is  venous  stenosis 
and  edematous  impregnation.  Some  investigators  have  found  the 
motor  ganglia  cells  of  the  ventral  horn  atrophically  vacuolized.  The 
nucleus  lies  often  asymmetrically,  pushed  to  one  side  by  a  large 
vacuole.  Rumpf  and  Luce  found  fresh  parenchyma  and  diffused 
medullary  sheath  degeneration  in  the  white  substance  of  the  spinal 
cord.  The  pathological-histological  changes  in  the  nervous  system 
will  be  further  described  under  the  subject  of  pellagra,  accompanied 
by  numerous  illustrations,  for  according  to  the  view  of  an  eminent 
neuro-pathologist,  Mott  (890),  these  changes  belong,  in  both  diseases, 
to  the  same  type;  however,  it  should  not  therefore  be  said  that 
both  diseases  must  be  etiologically  related. 

The  affected  muscles  show  all  signs  of  degeneration  and  atrophy. 
The  first  signs  of  this  consists  of  the  dimming  of  the  striations.  Fre- 
quently, the  muscle  fibers  become  waxy,  homogeneous  and  fragile. 
For  the  remaining  pathological  changes,  we  may  refer  to  what  has 
been  said  under  the  subject  of  avian  beriberi. 


BERIBERI  293 

Chemical  pathology — Blood 

The  blood-sugar  content  in  35  cases  of  infantile  beriberi  was 
determined  by  Suzuki  (891)  to  be  0.0808  per  cent.  This  decrease 
was  attributed  to  a  decreased  activity  of  the  adrenals,  and  is  different 
from  that  noted  in  avian  beriberi.  DeLangen,  Schut,  Wechuizen  and 
Alting  (892)  found  a  very  low  lipoid  and  fat  content  in  the  blood  in 
beriberi.  Arima  (893)  investigated  the  non-protein  nitrogen  in  28 
cases  and  found  it  increased. 

Cerebrospinal  fluid 

Suzuki  (894)  found  the  sugar  content  of  cerebrospinal  fluid  to  be 
normal. 

Urine 

Suzuki  (895)  found  the  amino-nitrogen  of  the  urine  markedly 
increased  in  the  cardiac  form  of  beriberi.  Pagniez  and  Vallery-Radot 
(896)  studied  the  elimination  of  sodium  chloride  on  a  diet  poor  in 
salt  in  cases  of  wet  beriberi.  The  elimination  was  very  large,  indi- 
cating a  previous  marked  retention.  In  the  wet  form  of  beriberi, 
as  in  nephritic  edema,  sodium  chloride  passes  the  kidney  only  with 
great  difficulty;  in  beriberi,  the  water  gathers  in  the  muscles,  while  in 
nephritic  edema,  the  fluid  collects  subcutaneously.  These  inves- 
tigators believed  that  the  development  of  wet  or  dry  beriberi  is 
determined  by  the  severity  of  the  kidney  lesions. 

In  confirmation  of  the  adrenaline  finding  in  pigeons,  Ono  (897) 
investigated  the  adrenals  microscopically  and  found  a  larger  quantity 
of  adrenaline,  as  compared  with  the  amount  in  other  pathological 
conditions. 

THERAPY   OF   BERIBERI 

General 

Vitamine  therapy  in  beriberi  is  at  present  so  well  established  that 
we  need  not  dwell  on  this  point  unnecessarily.  One  of  the  first 
investigations  was  that  of  Thompson  and  Simpson  (I.e.  60),  who  used 
yeast  for  this  purpose  with  good  results.  The  experiments  were 
thereupon  extended  by  Vedder  and  Williams  (I.e.  471)  to  include  a 
preparation  of  rice  polishings.  They  thought  that  they  had  obtained 
good  results  in  dry  beriberi  with  a  vitamine  preparation,  made 


294  THE    VITAMINES 

according  to  our  procedure,  while  in  wet  beriberi  it  was  ineffective. 
From  this,  it  was  concluded  that  for  the  therapy  of  various  types  of 
beriberi,  vitamines  of  various  sources  were  necessary.  This  observa- 
tion may  be  explained  by  the  severity  of  the  symptoms,  without 
having  recourse  to  such  a  hypothesis.  Good  results  with  vitamine 
preparations  were  obtained  by  Williams  and  Saleeby  (I.e.  472). 
Later  on,  Saleeby  (898)  used  autolyzed  yeast  in  adults  and  children. 
The  dose  used  was  15  to  40  cc.  three  times  daily  for  adults,  and  2  to  4 
cc.  every  3  hours  for  children,  greater  doses  being  of  no  advantage. 
Marked  results  were  noted  after  3  days,  but  the  treatment  was  con- 
tinued for  14  days  more.  In  infants,  the  results  were  noticeable  even 
sooner.  A  standardization  of  the  extract  of  rice  polishings  was 
recommended  by  tel  Rosario  and  Maranon  (899) .  In  the  late  war,  a 
yeast  preparation  (marmite)  was  used  in  the  English  army  with 
splendid  results  (Willcox,  I.e.  864).  Hepburn  (I.e.  845)  too  reported 
results  with  the  vitamine  therapy  in  Siam. 

Therapy  in  infantile  beriberi 

This  consists  either  of  a  change  of  milk,  or  of  the  administration  of 
vitamine.  Particularly  good  results  were  obtained  with  an  extract 
of  rice  polishings  (tiki-tiki).  According  to  Albert  (900),  the  symp- 
toms disappear  in  a  few  days,  sometimes  after  a  few  hours.  Cox 
(901)  remarked  particularly  on  the  decrease  in  the  number  of  cases 
of  infantile  beriberi  in  the  Philippines  since  the  introduction  of 
vitamine  therapy. 

It  need  hardly  be  said  that  poor  results  have  also  been  obtained 
by  the  use  of  vitamine  therapy.  What  we  said  about  avian  beriberi 
applies  here  also,  namely,  that  for  a  successful  vitamine  therapy  it 
is  necessary  throughout  that  the  anatomical  changes  should  not  have 
progressed  too  far. 

SHIP-BERIBEKI 

It  is  theoretically  quite  possible  that  vitamines  B  and  C  should 
both  be  lacking  in  the  diet,  and  in  this  way  give  rise  to  a  mixed  form 
of  scurvy  and  beriberi.  Such  mixed  forms  have  not  been  described 
with  great  certainty,  but  it  would  be  possible  to  regard  ship-beriberi 
as  such.  The  various  investigators  are  not  agreed  as  to  the  nature 
of  this  type  of  beriberi.  It  occurs  on  sailing  vessels,  when  the  supply 


BERIBERI  295 

of  fresh  vegetables  approaches  the  end.  Characteristic  symptoms 
are  numbness  of  the  extremities,  short  breathing,  and  finally  death 
from  heart  failure.  The  patients  recover  very  rapidly  when  they 
receive  fresh  provisions,  nervous  symptoms  being  noted  only  seldom; 
this  is  not  the  case  in  ordinary  beriberi.  If  the  crew  is  made  up  of 
representatives  of  rice-eating  people,  who  keep  to  their  dietary 
customs,  then  true  rice-beriberi,  and  not  ship-beriberi,  develops. 
Since  long  sea  voyages  are  now  of  rare  occurrence,  ship-beriberi  is 
rarely  met  with.  During  the  World  War,  the  German  raider 
Kronprinz  Wilhelm,  with  a  great  number  of  cases  of  ship-beriberi, 
was  interned  in  New  York  harbor.  The  crew  lived  on  frozen  meat, 
while  the  officers  received  a  daily  portion  of  fresh  vegetables  and  fruits 
in  addition,  and  therefore  did  not  contract  this  disease.  Since  fruits 
and  vegetables  contain  both  vitamines  B  and  C,  the  above  cases 
can  not  give  us  any  clue  as  to  the  true  nature  of  the  above  condition. 
Nocht  (902)  was  of  the  opinion  that  the  disease  was  not  identical 
with  rice-beriberi,  while  Vedder  (I.e.  838)  believed  that  it  was.  Hoist 
and  Frolich  (903),  on  the  contrary,  saw  therein  a  greater  resemblance 
to  scurvy,  although  Hoist  (904)  stated  later  that  on  Norwegian  ships 
the  crews,  which  ate  rye  bread,  were  free  of  this  disease.  However, 
if  the  rye  was  mixed  with  wheat,  the  disease  developed.  From  this, 
it  might  be  concluded  that  we  are  dealing  with  beriberi,  in  which 
view  Hoist  was  strengthened  by  the  results  of  his  pigeon  experiments. 
Ship-beriberi  is  apparently  a  mixed  syndrome,  differentiated  from 
scurvy  by  numbness  of  the  extremities,  and  from  beriberi  by  scor- 
butic gum  changes. 

RELATIONSHIP   BETWEEN   BERIBERI   AND   SCURVY 

As  we  have  already  seen  in  the  previous  chapter,  we  believe  that 
under  certain  conditions  mixed  avitaminoses  of  beriberi  and  scurvy 
may  arise,  though  in  practice,  from  a  lack  of  both  vitamines,  one 
condition  develops  in  preference  to  the  other.  It  is  nevertheless 
possible  that  scurvy  should  develop  on  an  exclusive  diet  of  rice. 
During  the  siege  of  Paris  in  1871,  if  the  observations  are  correct, 
such  cases  occurred  and  were  reported  by  Delpech  (905)  and  Bucquoy 
(906).  Garcia  (907)  described  a  mixed  form  of  scurvy  and  beriberi. 
Scherer  (908)  had  the  opportunity  of  observing  an  outbreak  of  scurvy 
in  German  Southwest  Africa,  in  which  there  was  a  death  rate  of  30 
per  cent.  The  disease  developed  because  of  an  exclusive  rice  diet 


296  THE    VITAMINES 

and  was  accompanied  by  beriberi  symptoms  (heart  dilatation).  In 
individual  cases,  anasarca,  serous  fluids  in  the  pericardium,  pleura  and 
peritoneum  were  noted — symptoms  which  belong  sometimes  to 
both  scurvy  and  beriberi.  Greig  (I.e.  880)  also  observed,  in  cases  of 
epidemic  dropsy,  scorbutic  symptoms,  with  bleeding  of  the  gums  in 
10  per  cent  of  the  cases.  Gouzien  (909)  described  an  interesting 
picture  of  a  disease  in  Hanoi.  At  first  beriberi  developed;  if  white 
rice  was  then  replaced  by  the  hand-milled  variety,  the  entire  64 
patients  promptly  recovered,  only  to  develop  scurvy  somewhat  later. 
The  cases  of  scurvy  which  develop  on  decorticated  corn  in  South 
Africa  are  explained  with  some  difficulty.  Fleming,  Macaulay  and 
Clark  (910)  reported  thousands  of  such  cases  among  miners  in  South 
Rhodesia.  The  composition  of  the  diet  there  was  as  follows:  Mealie- 
meal  (a  corn  product  poor  in  vitamine  B) — 2  pounds  daily;  meat — 1 
pound  weekly;  beans — 2  pounds  weekly;  and  nuts  (monkey-nuts)— 
1J  to  2  pounds  weekly.  Orenstein  (911)  is  of  the  opinion  that  the 
differential  diagnosis  of  beriberi  and  scurvy  in  these  cases  was  not 
made  properly;  he  could,  however,  firmly  establish  the  point  that 
these  diseases  promptly  disappear  on  giving  fruit  and  vegetables. 
Darling  (912)  regarded  this  South  African  disease  as  scurvy,  whose 
picture  was  complicated  by  beriberi.  Among  the  pathological  find- 
ings, there  was  noted  hypertrophy  of  the  right  heart,  fatty  degenera- 
tion of  the  heart  musculature  and  the  vagus,  in  addition  to  accen- 
tuated reflexes.  In  a  later  contribution,  Darling  (913)  stated  that 
scurvy  at  the  Rand  mines  can  not  be  so  easily  cured  as  pure  scurvy, 
and  it  was  therefore  regarded  as  a  mixed  form. 

What  we  wish  especially  to  establish  by  these  statements  is  the 
fact  that  we  encounter  in  beriberi,  as  well  as  in  other  human  avitam- 
inoses,  a  pathological  condition  which  perhaps  can  not  be  regarded 
as  being  entirely  due  to  the  lack  of  a  single  vitamine. 


CHAPTER  II 

SCUEVY 

A  historical  survey  of  this  interesting  disease  is  given  by  Schroder 
(914),  Schelenz  (915),  Hirsch  (916)  and  Autran  (917);  in  a  new 
monograph  by  Hess  (918),  the  whole  field  of  scurvy  is  critically 
treated.  We  have'  already  touched  lightly  upon  the  develop- 
ment of  this  subject  and  have  spoken  of  the  accurate  conception 
of  the  nature  of  scurvy  by  physicians  in  the  eighteenth  century — 
Kramer  (I.e.  32),  Bachstrom  (I.e.  33),  Lind  (I.e.  34).  From  all  of 
these  data,  we  see  that  scurvy  has  been  known  for  hundreds  of  years, 
and  makes  it  appearance  particularly  on  long  sea  voyages,  polar 
expeditions,  and  in  time  of  war.  Cook  (919),  in  the  report  of  his 
expedition,  mentioned  the  foodstuffs  needed  for  the  prevention  of 
this  disease.  On  one  occasion,  raw  sea-lion  meat  was  used  for  this 
purpose;  on  a  second  voyage,  he  used  a  malt  infusion.  Based 
upon  these  and  similar  experiences,  the  English  navy  in  1804,  at 
the  suggestion  of  Sir  Gilbert  Blaine  (920),  introduced  the  daily  portion 
of  lime  juice,  whereupon,  according  to  Budd  (921),  the  number  of 
cases  decreased  markedly.  It  appears  certain  that  scurvy  was  prev- 
alent in  every  war,  including  the  last,  in  great  numbers;  however, 
in  times  of  peace  it  was  due  to  bad  harvests,  especially  of  potatoes. 
MacNab  (922)  saw  numerous  cases  in  the  English  army  at  Nassirabad 
in  Raj  put  ana  (India)  during  1833-1834.  The  disease  was  brought  to 
an  end  by  using  a  native  sour  plant,  Phyllanthus  emblica.  In  the 
Civil  War  in  the  United  States  (923),  there  occurred  30,714  cases  of 
scurvy,  because  of  the  use  of  dried  vegetables  in  winter.  In  the 
Franco-Prussian  War  of  1870-1871,  numerous  cases  were  observed 
during  the  siege  of  Paris,  described  by  Delpech  (I.e.  905),  Bucquoy 
I.e.  906)  and  other  physicians  (924).  The  disease  was  at  time  time 
prevalent  in  prisons  and  was  frequently  accompanied  by  edema.  In 
the  Russo-Japanese  War,  scurvy  was  observed  among  the  Russians 
and  was  described  in  detail  by  Sato  and  Nambu  (925)  and  Blau  (926) . 
Sir  W.  G.  MacPherson  (927)  saw  20,000  cases  during  the  siege  of 
Port  Arthur.  As  regards  the  disease  among  the  civilian  population, 
it  made  its  appearance  when  there  was  a  scarcity  of  potatoes  and 

297 


298  THE    VITAMINES 

fresh  vegetables,  especially  when  large  amounts  of  meat  were  not 
available.  Curran  (I.e.  781)  reported  an  outbreak  of  scurvy  in 
Swift  Hospital,  Dublin,  in  1847,  among  patients  eating  almost  a 
pound  of  meat  daily,  or  drinking  a  half  liter  of  milk,  in  addition  to 
their  usual  food.  Rae  (928)  reported  that  in  the  Hudson  Bay  Expe- 
dition, the  natives  did  not  develop  scurvy  because  they  ate  a  large 
amount  of  meat  per  day — the  men,  8  pounds;  the  women,  6  pounds, 
and  the  children,  2  pounds.  Lanceraux  (929)  reported  cases  in  the 
prisons  of  the  Department  of  the  Seine  arising  from  a  lack  of  fresh 
vegetables  and  potatoes.  In  Russia,  the  disease  has  been  prevalent 
almost  at  all  times,  and  one  of  these  occurrences  has  been  described 
by  Berthenson  (930).  Miiller  (931)  observed  numerous  cases  in 
Ntirnberg  early  in  the  winter  of  1911,  arising  as  a  result  of  a  poor 
harvest.  Taussig  (932)  believed  that  in  Austria,  also  before  the 
World  War,  scurvy  was  not  infrequently  noted,  especially  when  it 
was  difficult  to  obtain  potatoes.  The  importance  of  potatoes  as  a 
protective  against  scurvy  is  firmly  established,  and  this  is  also  shown 
by  the  newest  data.  In  England,  especially  in  1917,  cases  of  scurvy 
occurred  in  poorhouses  in  Glasgow  (Pickens,  933)  and  in  Newcastle 
(Harlan,  934),  which  could  be  attributed  to  the  above  cause  (lack  of 
potatoes).  In  a  health  report  of  the  city  of  Manchester  (935),  cases 
of  scurvy  were  described  in  the  spring  of  the  same  year.  An  article 
in  the  British  Medical  Journal  (936)  at  this  time  showed  the  serious 
situation,  and  the  danger  of  the  substitution  of  potatoes  in  the  diet 
by  rice  and  bread.  The  quantity  of  potatoes  necessary  to  protect 
against  scurvy,  according  to  the  Committee  of  the  English  Royal 
Society  (937),  is  about  one  pound  per  day.  Hess  (I.e.  819)  showed 
the  etiological  relationship  between  the  occurrence  of  the  disease  and 
the  amount  of  potatoes  allotted,  in  one  institution  where  200  cases 
were  involved.  Lind  (938)  describes  a  number  of  cases  in  a  hospital 
at  Kew,  Victoria  (Australia),  while  G.  R.  Hopkins  (939)  observed 
3000  cases  in  Aruba,  a  small  island  in  Dutch  Guiana,  which  had  a 
total  population  of  10,000  in  1915.  The  cause  of  the  disease  was  a 
total  crop  failure  during  the  period  1912-1914.  The  diet  of  the 
natives  consists  of  corn,  corn  meal,  salt  fish  and  a  small  amount  of 
meat,  mostly  dried  or  salted.  The  well-to-do  class,  which  imports 
fresh  vegetables  and  fruits,  were  free  from  this  disease.  With  the 
arrival  of  the  rainy  period,  and  the  consequent  availability  of  fresh 
vegetables,  the  disease  disappeared  completely. 


SCURVY  299 

During  the  World  War,  thousands  of  cases  were  observed,  and  it  is 
most  likely  that  no  single  army  was  entirely  immune.  Hehir  (I.e. 
867)  described  numerous  cases  among  the  Indian  troops,  in  Mesopo- 
tamia, who  refused  to  eat  meat  because  of  religious  scruples.  Turner 

(940)  states  that  30  to  50  per  cent  of  these  troops  developed  scurvy, 
and  Wilcox  quoted  the  following  figures: 

1916 11,445 

1917 2,197 

1918 826 

On  the  other  hand,  among  the  English  troops,  only  sporadic  cases 
were  seen.  The  situation  was  so  serious  that  a  corps  of  gardeners 

(941)  was  sent  to  Mesopotamia  by  the  English  government  to  plant 
vegetables  of  every  kind  and  to  distribute  seeds  which  could  be 
sprouted  when  the  need  developed.     L.  Hill  (942)   described  an 
occurrence  of  scurvy  among  English  troops  in  Scotland,  attributed 
to  excessive  cooking  of  vegetables.     The  disease  seemed  to  single  out 
particularly  those  men  who  were  engaged  in  hard  work. 

During  the  late  war,  the  disease  was  noted  in  Russia  by  Horschelmann 
(943)  and  among  the  Russian  prisoners  in  Turkestan,  by  Disque  (944). 
Borich  (945),  chief  surgeon  of  a  Red  Cross  Station  in  Russia,  person- 
ally observed  1343  cases.  Among  some  troops,  75  per  cent  of  the 
men  were  disabled  by  this  disease. 

In  France,  Harvier  (946)  reported  that  in  1917  95  per  cent  of  his 
800  troops  developed  scurvy,  while  at  the  same  time  the  disease  was 
discovered  in  other  detachments.  Benoit  (947)  saw  63  cases  out  of  a 
body  of  350  men,  but  only  of  a  transient  nature;  the  patients  recovered 
without  special  treatment.  Mouriquand  (948)  too  spoke  of  a  number 
of  scorbutic  cases  in  the  army. 

For  the  Italian  army,  we  have  the  reports  of  Ferrari  (949), 
Ramoino  (950)  Gingui  (951),  and  of  Vanutelli  (952),  who  observed 
200  cases  of  infectious  purpura  with  hemorrhagic  scurvy.  Vallardi 
(953),  in  Macedonia,  saw  cases  among  Italian  soldiers  who  mani- 
fested icterus  and  enlargement  of  the  glands. 

From  Germany  and  Austria,  we  have  many  reports  of  the  army 
medical  staff,  from  which  it  is  clear  that  many  of  the  cases  were 
diagnosed  (954)  as  purpura  or  rheumatism.  The  cases  of  scurvy 
during  the  war  were  so  numerous  that  special  stations  were  opened. 
Schreiber  (955)  observed  30  German  prisoners  suffering  from  scurvy, 


300  THE   VITAMINES 

which  was  at  first  diagnosed  as  purpural  rheumatism.  Speyer  (956) 
was  sent  by  the  German  government  to  Bulgaria  to  make  a  study 
of  scurvy.  Lobmeyer  (957)  observed  scurvy  in  the  Turkish  army,  but 
it  is  possible  that  some  of  these  cases  were  complicated  by  war  edema 
and  beriberi.  Other  cases  of  scurvy  manifested  during  the  war 
will  be  discussed  in  the  course  of  this  chapter.  Simultaneously 
with  scurvy  in  adults,  numerous  instances  of  infantile  scurvy  were 
noted  in  Central  Europe.  However,  it  is  no  longer  necessary  to 
differentiate  between  these  two  types. 

MODE   OF   DEVELOPMENT 

It  is  always  instructive  and  of  practical  importance  to  know  the 
exact  diets  that  lead  to  the  respective  avitaminoses.  With  regard 
to  scurvy,  such  data  are  at  hand.  Scherer's  report  (I.e.  908)  treat 
of  more  than  862  cases  of  scurvy  in  the  earlier  German  colonies, 
among  miners  and  railroad  laborers,  of  which  289  died.  The  diet 
there  consisted  of  a  kilo  of  white  rice  and  500  grams  of  meat  two  times 
weekly,  almost  without  fruits  and  vegetables.  The  first  symptoms 
appeared  in  5  to  6  weeks.  In  the  Bulletin  of  Tropical  Diseases 
(958)  there  is  a  description  of  a  dietary  which  led  to  numerous  cases 
of  scurvy  in  the  prison  at  Burma : 

QTQWIS.  QTdfflS. 

Hulled  rice 684      Spices 4 

Beans 114      Fish  paste 14 

Vegetables 285      Salt 7 

It  was  shown  that  an  addition  of  vegetables,  milk,  meat  or  fish 
had  little  effect  on  the  disease,  but  an  addition  of  sweet  potatoes 
was  effective.  Dyke  (959)  described  numerous  cases  among  South 
African  natives,  transported  to  France  for  war  work.  The  number 
of  patients  amounted  to  680  out  of  a  total  of  1700.  The  diet  con- 
sisted of  450  grams  frozen  or  canned  meat  and  225  grams  fresh 
vegetables.  In  addition,  corn  or  rice,  bread  and  margarine  were 
provided.  Vegetables  were  cooked  for  3  hours,  which  was  obviously 
partly  responsible  for  the  outbreak  of  the  disease;  aside  from  this, 
Dyke  noted  that  the  natives  drank  3  liters  of  beer,  called  "Kaffir," 
prepared  in  Africa  from  germinated  kernels,  but  in  France  from 
ungerminated  corn.  This  beer  is  supposed  to  protect  the  natives 
from  scurvy  in  their  home  land.  During  the  war,  Comrie  (960) 


SCURVY  301 

was  with  the  English  troops  in  North  Russia;  in  the  prisons  there; 
he  observed  scurvy.  The  disease  began  after  4  to  7  months  on  a 
diet  consisting  of  313  grams  flour  or  zwieback;  250  grams  rice,  oat- 
meal, peas  or  beans;  205  grams  frozen  or  canned  meat  or  salted  her- 
ring; 50  grams  bacon  or  pork;  7  grams  tea;  28  grams  sugar;  21  grams 
salt  and  14  grams  preserved  lime  juice.  Of  the  antiscorbutics 
investigated,  the  order  of  activity  was  as  follows :  Sour  milk,  fresh 
meat,  fresh  lemon  juice,  germinated  peas,  canned  fruit  and  germi- 
nated beans.  Stevenson  (961)  believes  that  in  adults  the  disease 
develops  in  4  to  8  months,  and  that  200  grams  cooked  vegetables 
daily  may  prevent  the  occurrence  of  the  disease.  According  to  Chick 
and  Dalyell  (962),  prolonged  cooking  of  vegetables  is  frequently 
responsible  for  the  outbreak  of  scurvy.  Forty  such  cases  were 
observed  in  Pirquet's  children's  clinic  in  Vienna;  these  cases 
developed  eight  weeks  after  the  fresh  vegetables  were  dealt  out, 
somewhat  sparingly.  These  investigators  believed  also  that  rapid 
growth  on  a  diet  very  rich  in  calories  is  especially  conductive  to  the 
development  of  scurvy.  During  and  after  the  war,  a  number  of  cases 
of  scurvy  were  observed  among  older  children,  which  was  very  seldom 
the  case  in  times  of  peace.  Tobler  (963)  commented  in  particular 
upon  this,  mentioning  more  than  200  such  cases.  Erich  Miiller 
(964)  describes  a  number  of  these  cases  in  the  orphan  asylum 
(Frederick  the  Great)  in  Rummelsburg  near  Berlin,  which  were 
attributed  to  the  use  of  dried  vegetables.  Weill  and  Dufourt  (965) 
noted  it  in  children  between  2J  and  6  years  old  in  a  zone  formerly 
occupied  by  German  troops. 

Regarding  infantile  scurvy,  "Barlow's  disease"  (cf .  reviews  by 
Morse,  966),  this  condition  occurs  almost  exclusively  in  artificially 
fed  children.  There  are,  however,  a  few  investigations,  for  example, 
tfiat  of  Netter  (957),  who  described  the  disease  in  breast-fed  infants; 
however,  from  a  critical  survey  of  these  cases,  Hess  (I.e.  918)  con- 
cluded that  these  data  were  uncertain,  although  theroretically  not 
impossible.  The  cause  of  the  disease  is  to  be  sought  in  the  heating 
of  the  milk  (pasteurization  or  sterilization).  The  feeding  of  artificial 
milk  products  such  as  condensed,  homogenized  or  preserved  milk, 
or  of  children's  proprietary  foods  as  chief  diet  (Cheadle  and  Poynton, 
968)  brings  about  the  disease.  After  the  introduction  of  the  soxhlet 
apparatus,  the  milk  was  not  infrequently  warmed  for  45  minutes 
and  longer.  The  first  extensive  investigation  of  this  disease  we  owe 


302  THE    VITAMINES 

to  Sir  Thomas  Barlow  (I.e.  35).  He  concluded  that  this  type  is 
identical  with  scurvy  in  adults,  as  did  also  Hoist  and  Frolich  (I.e. 
903)  and  Looser  (969). 

Through  the  work  of  Neumann  (970),  we  recognized  the  causative 
relationship  between  heating  the  milk  and  the  appearance  of  infantile 
scurvy.  He  observed  the  development  of  scurvy  on  a  milk  which  was 
first  pasteurized  at  the  dairy  farm,  and  on  the  next  day,  before 
consumption,  heated  in  a  soxhlet  for  10  to  15  minutes.  Neumann's 
conclusions  were  confirmed  by  Heubner  (971)  and  by  A.  Meyer 
(972).  Brachi  and  Carr  (973),  in  England,  reported  some  cases  of 
of  scurvy  on  cooked  milk.  In  1898  the  American  Pediatric  Society 
(974)  issued  a  report  on  356  cases,  all  on  artificial  feeding.  Up  to 
1908,  Cheadle  and  Poynton  (I.e.  968)  collected  80  such  cases. 

More  recently,  Hess  has  occupied  himself,  together  with  his  co- 
workers,  with  the  subject  of  infantile  scurvy.  Hess  and  Fish  (975) 
found  that  scurvy  in  children  may  be  brought  about  by  feeding  milk 
heated  for  30  minutes  at  145°F.  The  condition  may  be  prevented 
by  the  addition  of  antiscorbutics.  Sittler  (976)  stated  that  milk, 
warmed  for  5  minutes  on  the  water  bath,  does  not  cause  scurvy. 
Comby  (977)  described  41  cases,  on  proprietary  foods,  among  which 
there  were  several  sent  in  with  an  incorrect  diagnosis.  Epstein 
(978)  reported  a  number  of  cases  in  Prague  during  the  war,  at  a  time 
when  oranges  and  fresh  fruits  were  scarce ;  the  milk  was  evidently 
heated  several  times  before  using. 

We  must  also  consider  the  flour  diet  as  an  etiological  factor. 
Bendix  (979)  described  a  case,  on  cow's  milk  diluted  with  Kufeke's 
flour  food,  which  was  cured  by  raw  milk.  Even  if  the  carbohydrates, 
as  such,  do  not  hasten  the  symptoms,  they  act  unfavorably  because 
of  the  milk  dilution.  Hess  and  Unger  (I.e.  656)  have  shown  that 
malt  soup,  prepared  with  the  addition  of  alkali,  is  productive  of 
scurvy,  while  Gerstenberger  (I.e.  829),  under  certain  conditions, 
could  effect  a  cure  with  it.  Such  cures,  however,  may  also  be 
attributed  to  variations  in  the  quality  of  the  milk.  Fordyce  (980) 
believes  that  the  disease  can  be  attributed  to  the  use  of  proprietary 
foods;  he  rarely  saw  the  disease  in  children  less  than  six  months 
and  more  than  one  year  old.  Apart  from  the  facts  touched  upon 
here,  the  points  to  be  considered  in  infantile  scurvy  are  those  that 
have  been  discussed  in  the  description  of  the  nutritive  value  of 
milk. 


SCURVY  303 

SYMPTOMATOLOGY  AND  PROGRESS  OF  SCURVY 

The  primary  stage  in  scurvy  is  characterized  by  a  peculiar  pale- 
ness of  the  skin,  apathy,  melancholy,  muscle  weakness  and  slight 
dyspnea.  The  skin  becomes  yellowish,  dry  and  scaly,  and  is  covered, 
especially  on  the  lower  extremities,  with  petechiae  and  larger  livid 
spots,  due  to  subcutaneous  hemorrhages.  In  addition,  there  are 
frequently  pains  in  the  lumbar  region  and  the  legs,  often  re- 
garded as  rheumatic.  Changes  in  the  skin  are  frequently  noted; 
thus  Taussig  (I.e.  932)  reports  an  exanthema  of  the  legs.  Aschoff 
and  Koch  (981)  describe  small  hemorrhages  near  the  hair  papillae 
(piquete  scorbutique),  and  an  exanthema  localized  in  certain  parts 
of  the  body,  mostly  observed  in  the  scorbutic  cases  during  the  war 
in  Rumania  and  in  the  Carpathians.  Similar  skin  changes  were 
described  by  Rheindorf  (982).  Wiltshire  (983)  saw  among  the  3000 
cases  of  scurvy  in  the  Serbian  army  numerous  skin  symptoms,  which 
he  called  "hyperkeratosis,"  a  condition  occurring  in  87  per  cent 
of  his  cases;  later  he  noted  an  exanthema  of  the  petechial  type, 
localized  in  parts  of  the  body,  already  mentioned  by  Aschoff  and 
Koch.  Wiltshire  also  observed  conical  swellings  around  the  hair 
follicles,  v.  Niedner  (984)  observed  that  infectious  exanthema 
frequently  assumes  a  hemorrhagic  appearance  in  the  presence  of 
scurvy.  According  to  the  paper  of  Sato  and  Nambu  (I.e.  925),  the 
skin,  as  well  as  the  subcutaneous  tissue,  is  often  edematous. 
According  to  Wassermann  (985),  subcutaneous,  diffuse,  painful  swell- 
ings appear  on  the  thighs;  Speyer  (I.e.  956)  occasionally  noted 
abcesses  on  the  calves.  Swellings  in  the  knee  caps  also  appear. 
The  gums  become  dark  red,  soft  and  swollen,  especially  around 
carious  teeth,  although  this  characteristic  sign  may  be  entirely 
lacking.  Sometimes  the  gums  swell  so  that  the  teeth  are  entirely 
covered.  Zlocisti  (986)  described  an  ulcerous,  gangrenous  sto- 
matitis. Aschoff  and  Koch  noted  a  falling  out  of  the  teeth  with 
resultant  hemorrhages. 

In  later  stages,  the  patients  suffer  very  much  from  dyspnea  and 
palpitation;  they  faint  easily,  and  heart  weakness  progresses.  The 
sufferers  becomes  markedly  emaciated,  the  muscles  become  atrophic, 
and  contractions  are  seen  at  times.  Scherer  (I.e.  908)  described  a 
tip-toe  position,  which  he  believed  to  be  the  result  of  hemorrhage; 
Aschoff  and  Koch  described  a  characteristic  position  which  they 


304  THE   VITAMINES 

called  "Seiltaenzerstellung."  Richter  (987)  speaks  of  tetanus  symp- 
toms in  scurvy.  It  is  not  quite  clear  whether  these  symptoms 
belong  to  the  picture  of  scurvy  or  whether  caused  by  complications, 
for  example,  with  beriberi. 


FIG.  60.  SKIN  EXANTHEMA  IN  SCURVY  (AscnoFF-Kocii) 

The  pulse  is  rapid;  the  left  and  right  ventricles  are  frequently 
greatly  enlarged.  Ascites  fluid,  dystrophic  edema,  subperiosteal 
hemorrhages,  bloody-serous  fluid  in  the  pericardium,  pleura,  or  in 
any  of  the  large  joints — all  belong  to  the  picture  of  scurvy.  In 


SCURVY  305 

this  stage,  we  meet  with  persistent  diarrhea  with  bloody-serous 
stools.  Sometimes  there  is  icterus,  as  was  found  by  Scherer  (I.e. 
908)  and  by  Urizio  (988).  Death  results  with  complications  of  an 
infectious  nature;  although  it  may  also  be  due  to  heart  failure. 

Complication  with  hemeralopia  (night  blindness)  was  noted  by 
Zak  (989)  among  Russian  prisoners.     Hift  (990)  was  able  to  avoid 


FIG.  61.  "SEILTANZER"  POSITION  IN  SCURVY  (AscnoFF-Kocn) 

these  complications  by  means  of  liver  therapy,  which  shows,  perhaps, 
that  the  condition  of  scurvy  in  these  cases  was  complicated  by  still 
another  avitaminosis.  O'Shea  (991)  saw  300  cases  of  hemeralopia 
among  English  troops,  in  22  cases  of  which  the  fundus  of  the  eye 
was  examined.  In  three  of  these  cases,  the  only  change  was  a  definite 
paleness.  Changes  in  the  retina  in  scurvy  were  investigated  by 
Kitamura  (992). 


306  THE    VITAMINES 

Infantile  scurvy 

Infantile  scurvy  develops  between  the  6th  and  18th  month,  seldom 
earlier  or  later,  and  Hess  (I.e.  918)  differentiated  between  the  acute, 
sub-acute  and  latent  forms ;  this  view  was  accepted  by  Miller  (993) . 
The  subacute  form  is  not  very  well  defined  but  growth  ceases.  Aside 
from  a  somewhat  abnormal  color  of  the  skin  and  an  occasional  edema 
of  the  upper  eyelids,  no  further  pathological  signs  are  noted.  The 
child  is  capricious,  and  the  lower  limbs  are  somewhat  sensitive  to 
touch.  The  knee  reflexes  are  almost  always  markedly  accentuated. 
All  of  these  symptoms  are  promptly  overcome  on  the  addition  of 
antiscorbutics,  whereupon  growth  is  resumed,  according  to  Hess 
(I.e.  461). 

The  latent  form  is  even  less  pronounced,  and  the  children  recover 
in  an  extraordinary  manner  after  the  administration  of  antiscor- 
butics. According  to  Hess,  the  latent  form  occurs  especially  in 
cities,  where  pasteurized  milk  is  sold.  It  does  not  need  to  be 
particularly  exphasized  that  these  less  important  forms  of  infantile 
scurvy  occur  more  frequently  than  do  the  acute  forms.  Franchetti 
(994)  described  a  chronic  case  of  scurvy  in  an  older  child,  with 
bone  dystrophy,  painful  paraplegia,  cachexia,  anemia,  absolute 
anorexia,  but  without  hemorrhages. 

Muscle  weakness,  anemia,  and  anorexia,  are  the  initial  symptoms 
noted.  Pain  develops  in  the  legs;  the  child  holds  them  motionless 
and  cries  out  when  they  are  moved.  This  behavior  resembles  pseudo 
paraplegia,  according  to  the  statement  of  Comby  (995).  Swellings 
are  frequently  noted,  sometimes  only  on  one  extremity,  of  a 
periosteal  nature,  especially  of  the  tibia.  Besides  this,  swelling  of 
the  muscles  was  noted,  caused  by  hemorrhages.  The  skin  on  the 
arches  of  the  feet  is  frequently  edematous,  while  the  skin  over  the 
swelling  appears  pale.  If  teeth'  are  present,  the  characteristic  gum 
changes  occur.  The  spongy,  bleeding  swellings  become  ulcerous 
in  some  cases  and  the  teeth  may  loosen  and  fall  out.  Oliguria, 
described  by  Gerstenberger  (996),  is  quite  characteristic  of  this 
condition,  and  hematuria  less  frequently  so.  The  temperature 
remains  normal,  occasionally  going  up  to  39°C.,  following  blood 
resorption  after  hemorrhages  (Barlow  fever).  When  no  thera- 
peutic measures  are  applied,  death  may  ensue  as  a  result  of  heart 
failure  or  hemorrhages.  Initial  phases  of  scurvy  were  described  by 


SCURVY  307 

Cozzolino  (997).     He  recommended  the  frequent  inspection  of  the 
gums  for  the  purpose  of  the  speedy  recognition  of  the  disease. 

A  sort  of  eczema  was  noted  by  Hess  (I.e.  918)  in  8  cases.  In  one 
instance  a  symmetrical  erythema  was  observed,  resembling  pellagra 
erythema  very  strongly.  Proptosis  of  the  eyeball,  following 
hemorrhages,  was  observed  by  Cheadle  and  Poynton  (I.e.  968)  as 
well  as  by  Zilva  and  Still  (I.e.  456).  It  occurs  usually  in  the  left  eye. 
Schodel  and  Naumwerk  (998)  observed  enlargement  of  the  right 
heart;  Hess  (999)  noted  the  quite  common  occurrence  of  polypnea 
and  tachycardia,  apparently  because  of  lesions  in  the  pneumogas- 
tricus.  Erdheim  (1000)  also  described  heart  lesions  in  Barlow's 
disease. 

DIAGNOSIS 

From  the  foregoing,  the  clinical  picture  of  scurvy  is  quite  obvious 
and  becomes  even  more  simple  as  a  result  of  the  newer  possible 
differential  diagnosis,  by  the  administration  of  vitamine  C.  Changes 
in  the  complexion,  follicular  changes  in  the  skin,  petechiae,  sub- 
cutaneous hemorrhages,  hemeralopia,  changes  in  the  gums,  feeling 
of  fatigue,  shortness  of  breath,  rapid  pulse  (about  140) — all  these 
are  specially  characteristic  of  scurvy.  Some  times  it  may  be  confused 
with  rheumatism  and  perhaps  with  hemorrhagic  diathesis.  In 
order  to  differentiate  here,  the  anamnesis  is  of  value  as  is  also  the 
result  of  vitamine  therapy.  Occasionally  the  hematological  investi- 
gation may  be  used  for  the  diagnosis,  which,  however,  according  to 
Rosin  (1001),  shows  an  unspecific  picture  of  anemia.  The  so-called 
"tibialgia"  of  v.  Schrotter  was  designated  as  scurvy  by  Labor 
(1002). 

In  the  diagnosis  of  infantile  scurvy,  the  "white  line,"  described 
by  Frankel  (1003)  in  the  X-ray  of  the  bones,  has  been  regarded  as 
characteristic.  It  is  a  shadow  which  is  apparent  at  the  epiphyseal 
ends  of  the  long  bones.  This  indication  was  observed  by  Brown 
(1004)  and  by  many  others,  although  Hess  (I.e.  918)  did  not  believe 
that  it  was  significant  in  the  diagnosis.  He  observed  this  shadow 
also  in  cases  of  scurvy,  which  had  been  cured  for  a  long  time.  Hess 
and  linger  (I.e.  759)  have  introduced  a  capillary  resistance  reaction 
for  the  diagnosis  of  scurvy,  which  consists  in  the  compression  of  the 
arm  for  three  minutes  with  a  tourniquet,  whereupon  petechial  spots 
appear  if  scurvy  is  present. 


308  THE    VITAMINES 

Hematology 

A  number  of  investigators  have  recorded  their  findings  as  to  the 
blood  picture,  but  it  does  not  appear  that  specific  indications  were 
discovered.  On  this  subject,  there  is  the  work  of  Labor  (1005), 
Leitner  (1006),  Brandt  (1007),  Wassermann  (1008)  and  Benoit 
(1009);  the  last  investigator  reported  findings  in  63  adults.  Hess 
and  Fish  (I.e.  975)  made  hematological  studies  in  Barlow's  disease, 
including  the  clotting-time  of  the  blood,  which  was  found  normal. 

As  regards  the  chemistry  of  the  blood,  the  sugar  content  was 
determined  in  some  cases  by  Roily  and  Oppermann  (1010)  as  well 
as  Schumm  (1011),  and  found  normal.  Hess  and  Killian  (1012) 
determined  the  calcium,  sugar  and  urea  content,  as  well  as  the  dia- 
static  and  the  carbon  dioxide  combining  power  of  scorbutic  blood. 
They  found  a  moderate  acidosis,  and  a  smaller  calcium  content 
(different  from  rickets) ;  as  for  the  rest,  the  values  were  normal. 

Metabolism  experiments 

Bauman  and  Howard  (1013)  investigated  the  inorganic  metab- 
olism in  one  case  of  adult  scurvy.  The  chlorine  and  sodium  balances 
were  negative  and  were  not  much  more  favorably  affected  by  the 
administration  of  antiscorbutics.  The  sulphur  metabolism  in  adults 
was  found  abnormal  by  Labbe,  Haguinea  and  Nepreux  (1014). 
The  case  investigated  was,  perhaps,  in  the  convalescent  stage  and 
was  marked  by  a  large  urea  output,  indicative,  possibly,  of  the 
disappearance  of  edema.  Experiments  in  an  older  child  were  carried 
out  by  Lust  (1015),  and  especially  by  Lust  and  Klocman  (1016). 
The  child  showed  a  retention  of  all  inorganic  constituents,  which 
were  subsequently  eliminated  after  a  cure;  this  applies,  in  particular, 
to  calcium.  These  data,  which  seemed  to  be  somewhat  uncertain 
at  first,  appear  now  to  be  corroborated.  As  for  the  calcium  metab- 
olism, the  above  results  were  confirmed  by  Moll  (1017).  Frank 
(1018),  experimenting  with  two  cases,  found  a  calcium  retention  in 
the  acute  stage;  during  convalescence,  this  calcium  was  again 
eliminated. 

Pathology  of  scurvy 

A  splendid  monograph  on  this  subject  is  that  of  Aschoff  and  Koch 
(I.e.  981)  as  well  as  of  v.  Samson-Himmelstiern  (cited  by  Aschoff 
and  Koch,  I.e.  981);  with  the  exception  of  severe  cases,  only  well 


SCURVY  309 

nourished  corpses  came  to  necropsy.  The  subcutaneous  tissue, 
especially  of  the  extremities,  is  infiltrated  with  bloody-serous  fluid. 
Widespread  blood  extravasations,  old  and  new,  are  noted.  The 
adductor  side  of  the  legs  shows  more  hemorrhages  than  does  the 
abductor;  hemorrhages  in  the  muscles  themselves  are  rare.  Hemor- 
rhages occur  also  in  the  periosteum,  especially  in  the  tibia,  and  in 
younger  individuals  in  the  epiphyseal  ends  of  the  long  bones  and 
at  the  cartilage  ends  of  the  ribs.  Bone  investigations  in  adults 
were  made  infrequently  but  it  appears  that  here,  too,  fractures 
are  noted.  Rosary  is  often  observed,  as  in  rickets.  Enlargement 
of  the  right  and  left  ventricles,  with  a  possible  fatty  degenera- 
tion of  the  heart  muscles,  was  also  described.  Most  of  the  organs 
were  examined  microscopically  by  Aschoff  and  Koch.  In  the  bones, 
osteoporosis  and  a  disappearance  of  osteoblasts  were  found;  nothing 
of  importance  was  seen  in  the  endocrine  gland's.  Suprarenals  showed 
only  a  greater  lipoid  content.  Feigenbaum  (1019)  found  hem- 
orrhages in  the  spinal  cord;  Scherer  (I.e.  908)  found  hemorrhages 
in  the  dura  and  pia  as  well  as  rank  growth  of  connective  tissue  in 
the  liver. 

Ecchymoses  occur  frequently  in  the  pleura,  less  often  in  the  peri- 
toneum, and  in  severe  cases  in  the  lungs.  Bloody-serous  fluids  in 
the  pericardium  and  pleura — less  frequently  ascites  fluids — were 
found  by  Johnson  Smith  (1020). 

Barlow's  disease 

The  pathological  findings  were  described  by  Frankel  (1021),  the 
bones  in  the  infantile  form  having  been  frequently  investigated.  The 
periosteum  is  filled  with  blood  and  is  thickened,  but  free  from  small 
cell  infiltration;  between  the  periosteum  and  the  bones  widespread 
hemorrhages  are  found.  At  the  diaphyseal  ends  of  the  long  bones, 
the  bone  marrow  loses  its  lymphoid  character  and  forms  reticular 
tissue  poor  in  cells  and  blood  vessels  which,  in  stained  sections,  is 
characterized  by  light  color,  because  of  "Helles  Mark."  This 
degeneration  of  the  bone-marrow  is  specific  for  infantile  scurvy  and 
prevents  normal  ossification.  The  new  bone  formation  is  delayed 
or  completely  prevented,  and  the  bone  tissue  present  is  atrophied, 
particularly  in  the  ossification  zones.  In  rare  cases,  hemorrhages 
are  found  in  the  dura,  in  some  joints,  in  the  lungs,  spleen  and  kidneys. 
As  we  have  already  stated  in  discussing  guinea  pig  scurvy,  Hess  and 


310  THE    VITAMINES 

Unger  (I.e.  378)  showed  that  a  scorbutic  rosary  may  be  cured  by 
giving  vitamine  C.  It  may  seem  from  this  that  the  complications 
of  scurvy  by  rachitic  signs  frequently  mentioned  in  the  literature 
may  be  attributed  to  the  above  condition.  By  investigation  of  the 
bones,  however,  the  scorbutic  lesions  may  be  differentiated  from 
the  rachitic.  In  rickets,  we  find  a  broad  band  of  osteoid  tissue 
formed  at  the  epiphyseal  ends ;  there  is  a  broad  band  of  insufficiently 


FIG.  62.  SCORBUTIC  ROSARY 
"Scurvy"  Hess,  courtesy  of  J.  B.  Lippincott 

calcified  cartilage,  which  is  never  observed  in  scurvy.  In  scurvy, 
the  growth  of  the  osteoblasts  is  markedly  inhibited,  but  when  growth 
starts,  it  proceeds  in  a  normal  manner.  In  addition,  we  find  a 
difference  in  the  number  of  capillaries  in  the  cartilagenous  tissue  and 
in  the  marrow,  very  rare  in  scurvy,  but  much  more  frequent  in  rickets. 
Hess  (1022)  saw  also  a  degeneration  of  the  ventral  horns  in  the  lumbar 
region  of  the  spinal  cord. 


SCURVY  311 

Bahrdt  and  Edelstein  (1023)  analyzed  the  various  organs  in  infan- 
tile scurvy.  The  bone-marrow  was  poor  in  solids,  calcium  and  phos- 
phorus. This  was  also  true  of  the  muscles;  normal  figures  were 
obtained  for  the  other  organs. 

THERAPY 

Since  we  have  already  spoken  of  the  antiscorbutic  value  of  various 
foodstuffs,  we  need  not  go  further  into  the  matter  at  this  point. 
On  using  antiscorbutics,  the  symptoms  disappear,  in  most  cases, 
after  2  weeks.  Even  severe  symptoms,  such  as  hydropericardium 
and  hydrothorax  are  very  favorably  influenced.  Still,  when  the  cases 
have  progressed  too  far,  curative  results  may  not  be  obtained. 
Paget  (1024)  treated  133  cases  with  camel's  milk,  with  good  results. 
As  for  lemon  juice,  24  cc.  daily  appear  to  be  sufficient  for  prophylaxis. 
Germinated  peas  and  beans  are  also  used  in  practice.  They  were 
given  to  Serbian  soldiers,  with  good  results,  by  Wiltshire  (I.e.  806). 
The  treatment  of  postscorbutic  conditions  was  described  by  Schul- 
hof  (1025). 

As  for  the  therapy  in  Barlow's  disease,  it  consists  in  the  addition 
of  raw  or  concentrated  milk,  orange  peel  extracts,  canned  tomatoes 
(Hess  and  Unger,  I.e.  668),  and  natural  fruit  juices  (Delille,  1026). 
Hess  and  Unger  (I.e.  637)  reported  successful  intravenous  therapy 
with  neutralized  orange  juice.  Harden,  Zilva  and  Still  (1027)  have 
used  dry  lemon  juice  preparations  with  great  success. 

With  regard  to  blood  transfusion  therapy,  Hess  (I.e.  918)  did  not 
find  much  vitamine  C  in  blood  so  that  a  large  amount  of  blood  would 
have  to  be  used  for  curative  purposes.  Similarly,  Rueck's  (1028) 
results  did  not  appear  any  more  certain.  With  a  well  chosen 
antiscorbutic,  the  therapy  is  almost  magical.  In  24  to  48  hours, 
the  curative  effect  is  already  distinctly  obvious. 


CHAPTER  III 

RICKETS 

This  universally  important  disease  occurs  invariably  in  the  first 
or  second  year  as  a  result  of  a  deficient  diet.  In  fact,  during  the 
last  war,  the  disease  occurred  almost  in  all  ages,  so  that  the  boundary 
lines  between  the  real  rickets,  rachitis  tarda  and  osteomalacia  have 
almost  disappeared.  As  we  shall  see,  the  inadequate  diet  of  itself, 
even  under  the  best  hygienic  conditions,  suffices  to  develop  the 
disease;  the  role  of  other  factors  such  as  lack  of  air  and  light,  is  not 
ruled  out.  By  and  large,  the  number  of  cases  of  rickets  increases 
with  the  rejection  of  the  natural  modes  of  nutrition  and  the  adoption 
of  artificial  feeding  of  infants.  The  further  child  nutrition  strays 
from  the  normal  (breast  milk),  the  less  it  corresponds  to  the  age  of 
the  child,  and  the  more  severe  are  the  symptoms  of  rickets.  This 
disease  is  chronic  and  is  not  dangerous  to  life  of  itself,  but  it  does 
predispose  towards  infections,  especially  of  the  respiratory  apparatus. 

Rickets  occurs  everywhere,  especially  in  the  temperate  zone,  less 
in  the  north  (Iceland,  Greenland,  Norway),  rarer  in  the  tropics  and 
also  southern  Italy  and  Spain.  Rickets  is  seldom  noted  in  countries 
where  breast  feeding  is  prevalent,  as  a  rule,  but  increases  in  the  in- 
dustrial centers,  following  artificial  infant  feeding  and  lack  of  sunlight. 
A  racial  influence  is  not  noticeable;  for  example,  rickets  frequently 
develops  among  negroes  and  southern  Italians  in  America,  whereas 
in  their  native  countries  they  are  unaffected  (Cautley,  1029;  Strong- 
man and  Bowditch,  1029a) .  The  frequently  observed  racial  predis- 
position may  be  attributed  as  well  to  the  dietary  customs.  Con- 
densed milk  together  with  proprietary  foods,  as  well  as  a  one-sided 
carbohydrate  diet,  leads  almost  invariably  to  the  development  of 
rickets.  According  to  Dennett  (1030),  there  is  no  danger  of  rickets 
from  cooked  milk,  as  such.  However,  even  breast-milk  may  be  an 
insufficient  food,  especially  when  the  mother's  diet  is  inadequate. 
We  have  noted  an  analogy  to  this  in  other  avitaminoses,  particularly 
in  beriberi. 

It  is  evident  that  rickets  arises  when  the  diet  either  is  lacking 
in  a  certain  vitamine  indispensable  for  normal  metabolism,  or  contains 
it  in  insufficient  amounts.  We  shall  speak  of  other  complicating 

312 


RICKETS  313 

factors  in  discussing  the  etiology  of  this  disease.  The  missing  sub- 
stance is  found  in  good  breast-milk  and  in  cod  liver  oil,  but  not  in 
certain  milk  preparations  and  flours.  It  has  nothing  to  do  with  a 
lack  of  fat,  protein  or  calcium,  per  se,  as  has  been  often  stated,  but 
with  a  vitamine  which  we  call  "anfcirachitic  vitamine,"  either 
belonging  to  or  identical  with  the  type  of  A-vitamine.  In  what 
way  the  lack  of  vitamine  A  brings  on  all  of  the  known  symptoms  of 
rickets,  is  at  present  unknown,  and  we  find  ourselves  upon  the  same 
uncertain  foundation  characteristic  of  the  explanation  of  all  other 
avitaminoses. 

OCCURRENCES 

According  to  the  newly  found  facts,  it  is  unlikely  that  the  diseasa 
is  associated  with  any  definite  age,  although  it  seems  certain  that  it 
is  manifested  mostly  during  the  period  of  active  bone  growth. 
Rickets  occurs  very  frequently  and  we  need  mention  only  two  investi- 
tigations  to  illustrate  this.  Schmorl  (1031)  stated  that  on  post- 
mortem of  the  cadavers  of  children  under  4  years  old,  90  per  cent 
showed  signs  of  rickets.  Schwartz  (1032)  observed  among  4944 
cases,  15  per  cent  craniotabes  in  the  first  year;  35  per  cent  showed 
rosary  (of  which  13  per  cent  were  in  the  first  month).  Congenital 
rickets  too  was  observed  with  certainty  (Kassowitz,  1032a).  Mery 
and  Parturier  (1033)  saw  a  case  in  a  6-weeks  old  child.  Wieland 
(1034)  reported  on  this  too,  but  he  (1035)  did  not  believe  in  the  exis- 
tence of  congenital  rickets.  Sinclair  (1036)  and  Carr  (1036a),  held 
that  premature  children  contract  rickets  very  easily,  since  the  neces- 
sary reserve  material  has  not  yet  been  prepared.  Hamilton  (1037) 
has  described  craniotabes  in  the  second  month  among  premature 
babies.  Retterer  and  Fisch  (1038)  describe  a  case  of  congenital  mi- 
cromelia  which  they  regarded  as  rickets.  Since  rickets  may  occur  at 
any  age,  the  cases  of  hereditary  rickets  are  not  puzzling;  the  mother 
may  be  suffering  from  a  lack  of  vitamine  A  during  pregnancy,  when 
the  requirements  are  even  greater  than  normally.  Among  the  cases 
of  "osteogenesis  imperfecta,"  there  are,  to  all  appearances,  those  that 
may  be  classified  as  congenital  rickets.  On  the  other  hand,  some 
of  them  must  be  regarded  as  hereditary  osteoporosis,  with  a  poor  cal- 
cium assimilation.  Ostheimer  (1039)  described  a  condition  in  chil- 
dren, called  "fragilitas  ossium,"  resembling  osteoporosis  (lack  of  cal- 
cium), which  in  this  case  could  be  improved  not  by  a  calcium  or  cod 


314  THE   VITAMINES 

liver  oil  therapy,  but  by  changing  the  diet  of  the  mother.  He  believed 
that  this  disease  was  associated  with  a  disturbance  in  the  metabo- 
lism of  the  mother.  One  case  of  osteogenesis  imperfecta  has  been 
described  by  Bookman  (1040).  In  this  instance,  calcium  lactate  and 
cod  liver  oil  were  used  with  good  results,  but  it  is  not  quite  clear 
which  of  the  two  was  responsible  for  the  curative  effect.  This  child 
progressed  even  better  on  breast-milk.  Another  such  case  was  re- 
ported by  McClanahan  and  Willard  (1041);  the  child  was  born  with 
multiple  fractures.  Another  case  was  improved  with  cod  liver  oil, 
according  to  Schabad  (1042). 

A  logical  transitional  stage  is  the  rickets  in  breast-fed  children. 
Brade-Birks  (1043)  reported  such  a  case,  and  in  this  respect,  Hess 
and  Unger  (1044)  controlled  the  diet  of  negro  women  in  New  York 
in  a  district  where  the  children  suffered  markedly  from  rickets. 
The  diet  was  recognized  as  being  very  inadequate  from  the 
dietetic  viewpoint. 

The  deficient  nutrition  in  Europe  during  the  war,  especially  in 
the  central  European  countries,  brought  with  it  a  great  increase  in 
the  number  of  cases  of  rickets,  according  to  Japha  (1045).  The 
mortality  in  rachitic  age,  due  to  the  pulmonary  complications,  was 
greatly  increased  according  to  Engel  (1046).  Weiss  (1047)  reported 
that  90  per  cent  of  the  children  born  in  Vienna  after  1917  suffered 
from  rickets,  not  excepting  the  children  of  well-to-do  parents.  The 
disease  appeared  mostly  among  children  from  1  to  4  years  of  age. 
Adams  and  Hamilton  (1048)  came  to  the  same  conclusion  as  regards 
Germany.  Rickets  in  older  children  was  described  by  Schlee  (1049), 
Stetter  (1050)  and  Sauer  (1050a)  in  Germany,  and  by  Sutton  (1051) 
in  Australia;  the  literature  on  the  subject  was  reviewed  by  Frangen- 
heim  (1051a).  Schlatter's  disease,  which  consists  in  a  separation  of 
the  epiphyses  of  the  long  bones  and  other  similar  fractures,  was 
described  in  a  13-year  old  boy  by  W.  Miiller  (1052)  and  in  a  similar 
case  by  Paus  (1052a);  these  cases  were  regarded  as  late  rickets. 

The  effect  of  rickets  on  later  life  has  apparently  been  under- 
estimated. Apart  from  the  possibility  of  permanent  bone  deformity, 
as  reported  by  Park  and  Rowland  (1052b)  and  by  Brusa  (1052c), 
the  disease,  according  to  Engel  (1053),  exerts  an  influence  on  the 
growth  of  the  children,  perhaps  on  the  mental  development.  Accord- 
ing to  Karger  (1054),  these  children  are  less  active  and  mentally 
underdeveloped.  In  this  connection,  Leri  and  Beck  (1055)  inves- 


KICKETS  315 

tigated  the  French  soldiers,  and  noted  that  those  who  had  suffered 
from  rickets  in  childhood  showed,  aside  from  anatomical  changes, 
mental  deficiency  and  little  resistance  to  bodily  exertion. 

SYMPTOMATOLOGY   AND   DIAGNOSIS 

Rickets  is  primarily  a  disease  of  the  bones,  although  it  must  be 
regarded  as  a  general  disturbance  of  metabolism.  As  we  gain 
knowledge  of  this  condition,  the  latter  assertion  may  perhaps  become 
more  justified.  The  children  show  a  weak  muscle  tonus  and  perspire 
profusely,  particularly  on  the  scalp.  They  commence  to  walk 
rather  late;  the  fontanelles  remain  open  longer  than  usual  and  the 
course  of  the  disease  may  be  controlled  to  a  certain  extent  by  measure- 
ment of  the  opening.  Swellings  of  the  cartilage  ends  of  the  ribs 
(rosary)  are  easily  palpated,  and  sometimes  are  apparent  even  without 
this.  According  to  Juaristi  (1056)  the  eyes  in  rickets  are  more  round 
and  show  more  of  the  sclera,  associated  with  bone  changes  in  the 
fundus  of  the  eye;  the  penis  is  long  and  hangs  flaccid.  Evidently 
the  delayed  development  of  the  teeth  is  also  characteristic  of 
rickets;  the  enamel  is  often  lacking,  and  occasionally  is  covered  with 
a  greenish  deposit  which  has  been  emphasized  by  a  number  of  authors 
(1056a).  According  to  Lichtenstein  (1056b),  rachitic  children  exhibit 
an  abnormal  sense  of  taste.  Various  other  symptoms  associated 
with  slight  muscle  tonus  are  described.  Characteristic  skull  forma- 
tion of  rachitic  origin  is  also  observed,  and  as  soon  as  the  children 
commence  to  walk,  the  legs  bend  in  the  known  way,  because  of  the 
weight  of  the  body. 

The  diagnosis  presents  no  particular  difficulty;  the  differentiation 
from  scurvy  having  already  been  discussed.  The  differential  diag- 
nosis between  rickets  and  some  pseudo-rachitic  appearances,  such 
as  "osteoporosis,"  is  not  so  easily  made.  In  this  respect,  the  X-ray 
may  be  of  some  help,  in  addition  to  the  results  of  vitamine  therapy, 
as  compared  with  calcium  therapy. 

PATHOLOGICAL   ANATOMY  AND    CHEMICAL   PATHOLOGY   OF   RICKETS 

We  find,  in  the  work  of  v.  Recklinghausen  (1057)  a  careful  and 
splendid  treatise  of  this  subject.  The  chief  sign  of  rickets  is  the 
persistence  of  the  cartilage  in  the  uncalcified  condition.  The 


316 


THE    VITAMINES 


rachitic  bone  is  recognizable  by  its  abnormally  small  content  of 
alkaline  earths.  The  deposit  of  calcium  salts  is  prevented  in  that 
part  of  the  skeleton  which  takes  those  salts  during  normal  growth; 
in  fact,  according  to  Schmorl  (1058),  this  happens  in  the  entire  skele- 
ton. A  greatly  increased  resorption  of  the  already  calcified  bone 
is  not  characteristic  for  rickets;  rather  does  it  characterize  pseudo- 
rickets.  In  the  investigation  of  rickety  bones,  the  preponderance 
of  the  cartilage  over  the  bone  tissue  is  especially  marked;  for  in- 
stance, Dibbelt  (1059)  found,  in  normal  bone,  29.4  per  cent  cartilage 
and  in  rickety  bones,  71.3  per  cent.  According  to  Schmorl  (I.e.  1058), 
there  is  an  abnormal  formation  of  osteoid  tissue.  Important  changes 
in  the  bone-marrow  were  found  by  Marfan,  Bardouin  and  Feuille 
(1060).  Marrow  cells  were  discovered  in  places  where  they  do  not 
occur  normally.  These  cells  were  later  substituted  by  fibroid  cells 
with  outgrowth  of  cartilage  cells;  by  this  means,  the  disturbance  in 
the  function  of  the  osteoblasts  is  explained.  Hutinel  and  Tixier 
(1081)  have  confirmed  the  above  findings.  Kassowitz  (1062)  found, 
in  rapidly  growing  bones,  an  abundance  of  blood  in  the  epiphyseal 
ends,  causing  a  proliferation  of  the  cartilage  cells  and  disturbing 
the  normal  ossification.  Heubner  (1063)  and  Pommer  (1064),  how- 
ever, found  no  inflammation  but  only  a  seemingly  abnormal  prolifer- 
ation of  the  cartilage  tissue.  Ribbert  (1065)  also  noted  a  marked 
breaking  down  .of  cartilage  cells.  Besides  the  bone  findings,  muscular 
dystrophy  was  noted  in  severe  cases  by  Hagenbach  and  Burckhardt 
(1066),  Bing  (1067)  and  also  by  Banu  (1087a).  According  to  Mohr 
(1068),  the  central  nervous  system,  besides  the  muscles,  also  is 
affected.  Of  the  endocrine  glands,  du  Castel  (1069)  investigated 
the  thymus  and  found  it  hypertrophied.  Stolzner  (1070)  found  the 
suprarenals  small  with  a  slight  adrenaline  content,  while  Cattaneo 
(1071)  was  unable  to  confirm  this. 

As  for  the  chemical  findings,  Dibbelt  (I.e.  1059)  gave  the  following 
composition  for  normal  and  rachitic  bones: 


NORMAL 

RACHITIC 

Fat 

1  89 

7  50 

Ca3(PO4)->  

57.38 

15.11 

Mg-phosphate 

1  72 

0  78 

CaCO3  

8.95 

3.15 

Other  salts  

0.83 

2.20 

RICKETS  317 

Gassmann  (1072)  found  the  following: 


NORMAL 

RACHITIC 

CaO                               .                         .... 

24.0 

21.0 

P2O5 

33.0 

30  0 

CO,         .        ...   :    

3.0 

2.75 

MgO 

0.10 

0.53-0  74 

In  addition,  5  per  cent  of  organic  matter  was  found  in  rachitic 
bones.  Similar  analytical  results  were  obtained  by  Schabad  (1073). 
Simonini  (1074)  saw  a  relationship  between  the  calcium  content  of 
the  bones  and  the  teeth.  Rost  (1075)  found  the  water  content  of 
rachitic  bones  higher  than  in  the  normal;  the  ash  content  of  the 
ribs  and  the  vertebrae,  on  the  contrary,  was  20  to  69  per  cent  lower. 
Aschenheim  and  Kaumheimer  (1076)  found  the  calcium  content  of 
the  muscles  diminished  in  severe  cases.  The  calcium  content  of 
the  blood  was  found  to  vary  by  Aschenheim  (1077).  Denis  and 
Talbot  (1077a)  observed  a  low  calcium  content  in  the  serum.  How- 
land  and  Kramer  (1077b)  frequently  found  the  calcium  content 
of  the  blood  normal,  while  the  phosphorus  content  was  always  low. 
The  sugar  content  of  the  cerebrospinal  fluid  was  found  diminished 
by  Suzuki  (I.e.  894). 

METABOLISM 

In  rickets,  according  to  Dibbelt  (1078),  the  elimination  of  salts  in 
the  feces  is  increased,  but  decreased  in  the  urine,  and  sometimes 
entirely  absent.  If  a  cure  is  brought  about,  there  is  first  a  hyper- 
retention  of  calcium  salts  with  simultaneous  increased  elimination  in 
the  urine.  Since,  in  his  time,  the  etiology  was  associated  with  the 
amount  of  calcium  in  the  food,  Denton  (1079)  tried  to  determine  the 
calcium  requirement  in  children.  Aron  (1080)  tried  to  prove  that 
milk,  especially  mother's  milk,  contains  just  sufficient  calcium  to 
cover  the  minimal  needs.  The  analyses  of  mother's  milk  made  by 
Schabad  (1081)  were  for  the  same  purpose.  Orgler  (1082)  disputed 
the  results  of  Aron,  and  attributed  them  to  a  faulty  calculation  of  the 
calcium  requirement.  Cronheim  and  Erich  Miiller  (1083)  compared 
the  mineral  metabolism  of  normal  and  rachitic  children,  and  found 
no  marked  variations.  In  Aron's  cases,  the  characteristic  metabolic 
disturbance  had  perhaps  passed.  While  the  assumption  of  the  lack 


318 


THE    VITAMINES 


of  calcium  in  the  food  could  not  be  confirmed  in  general,  the  lesser 
utilization  of  calcium  in  rachitic  children  has  become  a  fact  which 
has  gained  support  from  the  numerous  reports  of  Schabad  (1084) 
and  others.  Schabad  (1085)  also  studied  the  phosphorus  metabolism 
in  various  stages  of  rickets  and  convalescence.  The  children  investi- 
gated were  of  different  ages,  both  breast  and  cow's  milk  being  used ; 
normal  children  were  studied  as  controls.  The  following  figures 
were  obtained: 


RELATIVE 

P^Ofi 

P9(~)e 

EXCESS  IN 

EXCRETION 
PER  KILO 
PER  DAY 

EXCRETION 
IN  PER  CENT 
OF  AMOUNT 

PARTITION 
OF  P2O6 
EXCRETION 

FECE8  IN 
RELATION  TO 

Ca3(P04)2 

DIET 

CONDITION 

GIVEN 

o 
a 

1 

0> 

fl 

1 

•s 

.s 

1 

o 

q 

H 

p 

H 

d 

p 

£ 

U 

gram 

gram 

( 

Normal  4-5  months 

0.023 

.0018 

65.3 

52.8  80.6 

19.4 

64.8 

Breast-milk  { 

Progr.  rickets, 

| 

i 

5-13  months  

0.034 

0.021 

122.2 

72.2 

60.8 

39.2 

52.8 

( 

Normal  3-6months 

0.214 

0.119 

70.7 

45.1 

65.2 

34.8 

77.8 

Cow's  milk  \ 

Progr.  rickets, 

( 

5-8|   months.... 

0.186 

0.077 

94.6 

39.3  39.3 

60.7 

14.1 

I 

Normal,  4-5  years. 

0.15 

0.102 

80.3 

51.5  64.4 

35.6 

22.0 

Mixed  diet  { 

Progr.  rickets, 

4  years 

0.091 

0.04 

102.1 

44.9 

44.1 

55.9 

39.8 

^ 

Developed  rickets   1   year,   5 

months  to  2  years,  7  months  .  .  . 

0.178 

0.069 

71.8 

27.5  38.2 

61.8 

(16.5) 

23.3 

Convalescent   2|  to  8  years  

0.103 

0.077 

65.5 

48.7  74.4 

25.6 

35.6 

(64) 

Very  recently,  an  important  series  of  metabolism  experiments  in 
rickets  was  reported  by  Schloss  (1085a)  and  also  by  Freise  and 
Rupprecht  (1085b). 

According  to  Flamini  (1086)  the  urinary  CaO  excretion  in  normal 
children  is  0.125  gram  per  day;  in  rickets,  0.05  gram  on  an  intake  of 
3.4  grams  per  day.  Peiser  (1087)  found  the  calcium  balance  negative. 
The  nitrogen  and  sulphur  metabolism  was  studied  in  rachitic  dwarfs 
by  Schwarz  (1088),  and  the  mineral  metabolism  in  late  rickets,  by 


RICKETS 


319 


Schabad  (1089).     Schabad's  findings  on  the  deficient  resorption  of 
calcium  were  corroborated  by  Dibbelt  (1090). 

The  study  of  the  influence  of  the  composition  of  the  diet  on  the 
utilization  of  calcium  gives  some  interesting  results.  The  effect  of 
protein  was  investigated  by  L.  F.  Meyer  (1091)  and  by  Tada  (1092). 
They  found  that  the  nitrogen  elimination  did  not  go  parallel  with  the 
calcium  output.  The  influence  of  fats  was  studied  by  Meyer  (I.e. 
1091),  Rothberg  (1093)  and  Orgler  (1094).  They  found,  surpris- 
ingly, that  the  addition  of  fats  (in  the  form  of  whole  milk)  acted 
unfavorably  on  the  calcium  utilization,  losses  occurring  in  the  feces 
in  the  form  of  soaps.  The  following  figures  were  obtained  on  com- 
paring the  effect  of  cod  liver  oil  and  butter  on  the  calcium  balance: 


WITHOUT 
COD  LIVER  OIL 

WITH 
COD  LIVKR  OIL 

INSUFFICIENT 
BUTTER  FAT 

EXCESS 
BUTTER  FAT 

gram 

gram 

gram 

gram 

1 

+0.060 

+0.175 

6 

+0.137 

-0.198 

+0.141 

7 

+0.038 

-0.034 

2 

-0.014 

+0.143 

8 

+0.043 

-0.120 

+0.519 

9 

+0.037 

-0.267 

3 

+0.073 

+0.303 

4 

-0.038 

-0.285 

+0.141 

+0.108 

5 

+0.067 

+0.465 

Hess  (1094a)  demonstrated  the  unfavorable  influence  of  butter. 
On  the  other  hand,  Telfer  (1094b)  found  that  fat,  whether  in  the 
form  of  cod  liver  oil  or  butter,  has  no  influence  on  the  calcium 
metabolism.  It  should  be  noted,  however,  that  the  duration  of  his 
experiment  was  too  short  (4-5  days) ;  besides,  the  child  under  investi- 
gation was  not  rachitic.  Hutchinson  (1095)  found  that  the  soap 
elimination  in  rachitic  stools  was  somewhat  decreased,  2.2  grams 
instead  of  2.5  grams  in  normal  children.  In  addition,  he  (1096) 
found  that  the  fat  loss  can  be  very  large  in  a  voluminous  stool,  giv- 
ing a  negative  balance,  under  certain  conditions,  of  0.6  gram  per 
day;  the  fat  sometimes  constitutes  one-third  of  the  total  volume 
of  stools. 

Holt,  Courtney  and  Fales  (1097)  demonstrated  a  large  loss  of 
fat  in  the  feces.  However,  the  poor  utilization  of  calcium  after 
the  addition  of  milk  fat  (butter)  could  not  be  corroborated  by  these 
investigators  (1098). 


320  THE    VITAMINES 

The  addition  of  carbohydrates  appears,  according  to  the  prevailing 
data,  to  influence  the  calcium  metabolism  favorably,  although  the 
results  were  not  constant.  Reports  on  this  subject  were  made  by 
Dibbelt  (1099),  Massaneck  (1100),  Tada  (I.e.  1092)  and  Rothberg 
(I.e.  1093).  Howland  and  Marriott  (1101)  have  investigated  the 
action  of  sugar  and  cereal  addition  and  found  it  favorable;  but  the 
effect  of  large  excess  of  carbohydrates  was  not  investigated. 

THERAPY    AND    THERAPEUTIC    EFFECT    ON   THE    METABOLISM. 

The  therapy  of  rickets  with  phospho-cod  liver  oil  was  introduced 
by  Kassowitz,  and  is  still  used  almost  in  the  original  form.  Turning 
to  the  findings  on  the  calcium  metabolism,  the  clinical  reports  on  the 
therapeutic  value  of  cod  liver  oil  are  not  particularly  uniform,  and  it 
may  be  concluded  that  perhaps  not  all  cod  liver  oils  possess  the  same 
therapeutic  value.  As  to  this,  we  have,  first  of  all,  the  prolific 
work  of  Schabad  (1102)  and  his  co-workers,  in  which  the  favorable 
effect  of  cod  liver  oil  is  demonstrated;  the  value  of  cod  liver  oil 
emulsions  was  also  tested.  In  a  later  investigation,  Schabad  (1103) 
studied  the  effect  of  the  Czerny-Keller  diet  on  rickets  and  found  that 
it  was  evidently  not  adequate,  since  the  addition  of  cod  liver  oil  still 
acted  favorably,  whereas  in  a  pure  milk  diet,  the  oil  was  unnecessary. 
Birk  (1104)  likewise  reported  on  the  results  of  phospho-cod  liver  oil 
therapy. 

It  is  only  questionable  whether  in  the  above  combination,  the 
phosphorus  addition  is  necessary.  Frank  and  Schloss  (1105)  believe 
that  between  the  two  preparations — cod  liver  oil  and  phospho-cod 
liver  oil — no  far-reaching  differences  exist;  however,  Schloss  (1106) 
recommended  a  simultaneous  addition  of  a  calcium  preparation. 
Grosser  (1107)  investigated  the  retention  of  organic  calcium  salts. 
Pereida  y  Elardi  (1108)  did  not  believe  in  the  favorable  effect  of  the 
calcium  addition,  since  a  physiologically  sufficient  amount  of  calcium 
is  supposed  to  be  present  in  the  diet.  Kurt  Meyer  (1109)  studied 
the  effect  of  cod  liver  oil  therapy  on  the  calcium  balance,  and  obtained 
good  results.  Tobler  (1110)  cured  four  cases  of  late  rickets  with 
phospho-cod  liver  oil,  and  similar  results  were  obtained  by  Ricklin 
(1111).  Phemister  (11 12)  was  of  the  opinion  that  phosphorus,  as  such, 
possesses  no  significance  in  the  therapy  of  rickets,  but  does  stimulate 
the  formation  of  osteoid  tissue.  Phemister,  Miller  and  Bonar 
(1112a)  believe  now  that  phosphorus,  per  se,  has  a  curative  action. 


RICKETS 


321 


However,  the  number  of  cases  investigated  is  insufficient,  besides 
which  there  is  the  possibility  of  a  spontaneous  cure  to  be  considered. 
As  a  result  of  their  rat  experiments,  Sherman  and  Pappenheimer 
(1112b)  and  Pappenheimer,  McCann,  Zucker  and  Hess  (1112c) 
are  inclined  to  the  belief  in  the  therapeutic  influence  of  phosphorus 
alone.  They  believed  they  could  produce  rickets  in  rats  on  a 
phosphorus-poor  diet,  and  then  effect  a  cure  by  the  addition  of 
phosphorus.  These  experiments  appeared  not  very  convincing  at 
first,  but  have  since  received  additional  experimental  proof.  Lienaux 
and  Huynen  (1113)  do  not  believe  in  the  therapeutic  influence  of 
calcium  in  rickets. 

In  opposition  to  the  above  more  or  less  definite  results,  there  are 
the  findings  of  Hess  and  Unge;  (1114)  on  negro  children  in  New  York. 
In  these  cases,  cod  liver  oil  was  used  without  any  phosphorus  addi- 
tion. In  the  negro  section,  about  90  per  cent  of  all  the  children  are 
rachitic;  the  mortality  is  about  314  per  1000,  in  which  tuberculosis, 
pneumonia,  and  whooping  cough  play  the  greatest  part,  it  being 
known  that  rickets  predisposes  towards  disease  of  the  respiratory 
organs.  Altogether,  40  children  were  given  cod  liver  oil,  while  16 
children  served  as  controls.  The  children  were  between  4  and  12 
months  old  and  the  complete  results  were  as  follows : 


AVERAGE  TOTAL, 
QUANTITY  OF 
COD  LIVER  OIL 

TIME  OP 
THERAPY 

NUMBER  OF 
CHILDREN 

RICKETS 

NO  RICKETS 

NO  RICKETS 

grams 

months 

per  cent 

1550 

6 

32 

2 

30 

93 

655 

6 

5 

1 

4 

80 

600 

4 

12 

5 

7 

58 

16 

15 

1 

6 

The  children  were  mostly  breast-fed  and  the  results  of  the  therapy, 
especially  in  cases  receiving  more  cod  liver  oil,  were  very  marked. 
These  investigators  (I.e.  1044)  studied  also  the  diet  of  the  mothers 
and  found  it  very  deficient.  The  results  of  Ferguson  (1115)  were 
about  the  same  as  the  above.  A  number  of  out  patient  cases  of 
rickets  were  treated  by  Mackay  (1115a):  Small  doses  of  cod  liver 
oil,  butter  or  cottonseed  oil  had  no  influence,  but  larger  doses  of 
cod  liver  oil  were  effective.  According  to  Aron  (1115b)  carrot 
extract  was  of  no  value  in  rickets. 


322 


THE    VITAMINES 


In  1912  Raczynski  (1115c)  formulated  a  very  interesting  theory, 
quite  in  accord  with  the  domestication  theory  of  Kassowitz.  He 
thought  rickets  developed  because  of  a  lack  of  sunlight,  a  view  which 
seems  to  have  been  completely  confirmed  by  the  newest  experimental 
investigations.  Huldschinsky  (1115d)  was  able  to  cure  rickets  by 
means  of  alpine  light,  since  the  X-ray  showed  a  marked  deposition 
of  calcium.  Mengert  (1115e)  used  the  quartz  lamp  for  prophylactic 
purposes  and  Erlacher  (1115f)  succeeded  by  this  means,  in  favor- 


FIG.  63.  X-RAY  DIAGNOSIS  OF  RICKETS  AND  THE  EFFECT  OF  COD  LIVEK  OIL 
THERAPY  SEEN  BY  THIS  METHOD 

Left,  May  22,  1920;  right,  October  31,  1920  (Hess) 

ably  influencing  46  cases  of  rickets,  without  any  change  in  diet. 
Hess  and  linger  (1115g)  used  sunlight  for  the  same  purpose,  with- 
out any  dietary  changes,  and  they  (1115h)  attributed  the  seasonal 
variations  in  the  incidence  of  rickets  to  the  effect  of  sunlight.  This 
explanation,  among  other  factors,  "sounds  quite  plausible. 

In  order  to  follow  the  course  of  the  therapeutic  measures,  it  is  not 
strictly  necessary  to  carry  out  metabolism  experiments.  It  may  be 
studied  by  means  of  the  X-ray,  a  method  utilized  by  a  number  of 


RICKETS  323 

investigators,  among  whom  should  be  mentioned  Phemister  (I.e. 
1112),  Rowland  and  Park  (1116),  and  Dufour  (1116a)  and  Hess. 
The  results  of  cod  liver  oil  therapy  are  apparent  in  animals  some- 
times after  only  two  days;  in  children,  after  about  three  weeks,  when 
the  deposition  of  calcium  may  actually  be  noted.  This  method  is 
coming  into  general  use,  and  will  very  likely  become  of  great  prac- 
tical importance. 

ETIOLOGY 

Although  we  accept  the  vitamine  etiology  in  rickets,  we  are  well 
aware  that  still  other  views  for  the  explanation  of  the  nature  of  this 
disease  have  been  advanced.  One  of  these  hypotheses,  the  causal 
relationship  with  the  endocrine  glands,  we  have  already  discussed  in 
detail  in  our  first  edition,  and  will,  therefore,  not  go  into  it  again  here. 
Since  the  function  of  the  vitamines,  especially  vitamine  A,  is  entirely 
unknown,  the  vitamine  hypothesis  in  rickets  does  not  exclude  the 
possible  role  of  the  glands  in  the  development  of  rickets.  The 
study  of  the  etiology  of  rickets  has  been  in  a  state  of  change  during 
the  past  year.  According  to  the  results  of  modern  experimental 
rickets  investigations,  this  disease  seems  to  be  associated  with  a 
number  of  factors  a  fact  which  might  assign  to  rickets  a  special 
place  in  pathology.  Three  such  factors  have  been  put  in  the  fore- 
ground— lack  of  vitamine  A,  lack  of  phosphorus  and  lack  of  sun- 
light. Each  of  these  factors  alone  is  able  to  cure  the  disease,  which 
develops  as  well  in  rats  and  dogs  as  in  man,  and  has  a  favorable 
influence  on  the  calcium  metabolism.  Similar  results  have  already 
been  obtained  with  children.  Nevertheless,  it  is  not  yet  clear  as  to 
just  how  these  three  factors  are  related  to  each  other.  They  give, 
at  all  events,  the  same  pathological  picture.  The  "domestication 
theory"  of  v.  Hansemann  (1117)  and  also  of  Kassowitz  (1118)  has 
recently  been  accentuated  by  a  number  of  English  investigators — 
Findlay  (1119),  Paton,  Findlay  and  Watson  (1120)  and  Ferguson 
(I.e.  1115),  and  by  the  modern  sunlight  hypothesis.  Dick  (1122) 
believes  that  rickets  is  a  disease  of  great  industrial  centers,  and  that 
it  does  not  occur  in  the  tropics,  Japan  or  China.  However,  this 
seems  to  have  little  foundation  in  fact.  Although  we  do  not  wish 
to  state,  at  this  time,  that  a  poor  hygienic  condition  has  nothing 
whatever  to  do  with  the  etiology  of  rickets,  we  believe,  nevertheless, 
that  it  can  not  be  the  only  cause.  For,  as  it  most  frequently  hap- 


324  THE   VITAMINES 

pens  in  practice,  the  unfavorable  hygienic  conditions  are  associated 
with  a  deficient  dietary.  Mann  (1123)  investigated  statistically 
1000  cases  of  rickets  in  London,  together  with  250  controls.  His 
results  speak  in  favor  of  the  dietetic  cause  of  rickets,  inasmuch  as 
he  paid  particular  attention  to  the  mode  of  nutrition.  A  critical 
survey  of  his  cases  convinced  him  that: 

44  per  cent  of  the  cases  suffered  from  a  lack  of  fat  and  an  excess  of  carbohy- 
drates in  the  diet. 

16  per  cent  were  breast  fed  by  a  poorly  nourished  mother. 

13  per  cent  occurred  in  families  in  straightened  circumstances. 

6  per  cent  exhibited  a  mis-proportion  between  the  dietary  constituents,  in 
favor  of  the  carbohydrates. 

Czerny  (1123a)  and  Siegert  (1123b)  allot  to  heredity  an  im- 
portant role  in  the  etiology  of  rickets.  That  the  cause  for  all  rachitic 
manifestations  lies  in  the  central  nervous  system,  was  assumed  by 
Pommer  (I.e.  1064),  and  later  by  Schabad  (1124),  but  this  assumption 
did  not  exclude  the  dietetic  factor.  Other  dietetic  theories  are  as 
follows : 

1.  Lack  of  fat  and  excess  of  carbohydrates;  Herter  (1124a);  Holt 
(1125);  Cheadle   (1126). 

2.  Lack  of  a  substance  of  an  unknown  nature  in  the  diet;  Hopkins 
(I.e.  23). 

3.  Lack  of  calcium  in  the  food;  Stolzner  (1127). 

4.  Excessively  rich  food;  Esser  (1128). 

5.  Lack  of  organic  phosphorus  combinations;  Schaumann  (I.e.  2). 

6.  Nutritional  disturbances  of  every  description,   acidosis  and 
excessive  nourishment;  Pritchard  (1129). 

7.  Lack  of  antirachitic  vitamine;  Casimir  Funk;  Mellanby. 

The  vitamine  etiology  of  rickets 

This  theory  was  first  suggested  by  us  and  was  attributed  to  the 
lack  of  a  specific  antirachitic  vitamine,  in  relation  to  which,  the  pres- 
ence of  such  a  vitamine  in  cod  liver  oil  has  been  assumed.  These 
conceptions  were  first  tested  experimentally  by  Edward  Mellanby  (I.e. 
95).  In  the  discussion  as  to  the  vitamine  requirements  of  dogs,  we 
have  already  touched  upon  his  work.  He  divided  the  foodstuffs 
used  by  him  into  two  groups — one,  protective  against  rickets,  the 
other,  not. 


RICKETS  325 

Not  protective  Protective 

Bread  Whole  Milk  (500  cc.  per  day) 

Oatmeal  Cod  liver  oil 

Rice  Butter 

Skim-milk  Lard 

Yeast  (10-20  grams  per  day)  Olive  oil 

Orange  juice  (5  cc.  per  day)  Arachis  oil 

Linseed  oil  Suet 

Babassu  oil  Cottonseed  oil 

Hydrogenated  fat  Meat 

Ca2(P  O4)3  Meat  extracts 
NaCl 
Milk  protein 

At  the  time  of  Mellanby's  first  publication,  the  view  was  largely 
held  that  vegetable  oils  and  fats  contain  no  vitamine  A  and  con- 
.sequently  the  above  table  was  not  so  significant  as  is  now  the  case. 
In  spite  of  some  differences,  hard  to  explain,  Mellanby  is  of  the 
opinion  that  vitamine  A  is  identical  with  the  antirachitic  vitamine. 
According  to  his  findings,  the  requirements  for  this  vitamine  are 
particularly  large  when  growth  is  most  rapid.  In  a  later  communica- 
tion, Mellanby  (1130)  stated  that  the  requirement  for  vitamine  A 
depends  very  much  upon  the  composition  of  the  diet.  The  high 
protein  content  of  a  diet  stimulates  the  total  metabolism;  the  children 
are  active  and,  therefore,  require  less  of  this  vitamine.  On  a  carbo- 
hydrate diet,  the  metabolism  is  sluggish  and  the  organism  requires 
more  of  this  vitamine.  However,  it  seems  more  logical  to  regard  the 
action  of  protein  as  "vitamine  sparing/'  judging  by  our  personal 
experience.  Besides  the  composition  of  the  diet,  age  also  plays  a  big 
part,  according  to  Mellanby  (1131).  Rickets  is  produced  in  older 
dogs  only  with  great  difficulty,  and  spontaneous  cures  of  ten-occur. 
That  this  is  not  quite  in  accord  with  human  pathology  is  evident  in 
rachitis  tarda  and  osteomalacia.  Therefore,  the  chapter  on  the 
etiology  of  rickets  can  not,  as  yet,  be  regarded  as  closed.  In  spite  of 
the  fruitful  investigations  of  Mellanby,  several  points  are  not  yet 
clear,  for  example,  the  prophylactic  influence  of  meat  extracts.  It  is 
not  impossible  that  Mellanby's  experiments  have  been  rendered 
somewhat  obscure  through  the  non-observance  of  the  light  factor 
which  will  have  to  be  considered  in  all  future  research  on  this  subject. 

Recently,  Noel  Paton  and  Watson  (1131a)  have  subjected  the 
researches  of  Mellanby  on  young  dogs  to  experimental  testing  and 


326  THE   VITAMINES 

concluded  that  rickets  in  dogs  is  not  dependent  upon  the  lack  of 
antirachitic  vitamine,  but  upon  the  energy  content  of  the  diet, 
exercise  and  hygiene.  However,  they  admit  that  dogs  on  skim- 
milk  develop  rickets  more  readily. 

Hess  and  Unger  (I.e.  96),  in  particular,  have  subjected  Mellanby's 
results  to  a  sharp  scrutiny.  They  maintained  five  children,  5  to 
12  months  old,  on  a  diet  composed  of  180  grams  Krystalak  (dried 
skim-milk  with  0.2  per  cent  fat),  30  grams  cane  sugar,  15  to  30  grams 
autolyzed  yeast,  15  cc.  orange  juice  and  30  grams  cottonseed  oil 
(cereals  for  older  children).  Some  of  these  children  were  kept  for 
several  months  on  this  diet,  and  as  they  grew  older  the  amount  of 
cereal  was  increased,  no  other  change  being  made.  At  the  time  of 
publication,  the  diet  was  thought  to  be  completely  free  of  vitamine 
A.  These  remarkable  experiments  showed  for  the  first  time  that 
vitamine  A  as  it  occurs  in  milk  plays  only  a  minor  part  in  the  develop- 
ment of  rickets.  This  finding,  which  contradicts  somewhat  the 
results  obtained  by  Mellanby  in  his  experiments  on  dogs,  was  not 
generally  accepted.  Hopkins  (I.e.  595)  held  it  against  Hess'  experi- 
ments that  the  above  diet  was  not  entirely  free  from  vitamine  A, 
since  skim-milk  may  still  contain  this  substance;  cottonseed-oil  too 
is  not  free  from  this  vitamine.  Although  these  objections  are 
partially  justified,  we  can  not  doubt  that  the  dietary  used  by  Hess 
and  Unger  was  indeed  very  poor  in  vitamine  A.  The  favorable 
result  obtained  with  this  diet,  extremely  poor  in  vitamine  A,  suggested 
that  the  diet  per  se  is  not  the  only  causative  factor  in  the  etiology  of 
rickets,  and  that  there  are  still  other  factors  to  be  taken  into  consider- 
ation. Should  they  be  so  interpreted,  that  rickets  has  nothing  to  do 
with  vitamine  A,  as  Hess  and  Unger  naturally  concluded  at  first? 
On  the  contrary,  these  important  experiments  show  that  under 
certain  conditions,  the  vitamine  A  requirements  of  children  may  be 
very  slight,  namely,  when  the  composition  of  the  diet  as  regards 
protein,  salts,  vitamines  B  and  C  leaves  nothing  to  be  desired.  The 
individual  differences  in  children  on  the  same  diet  may  perhaps  be 
attributed  to  unequal  utilization  of  dietary  constituents,  a  point 
which  has  not  been  allowed  for  in  the  experiments  of  Hess  and  Unger, 
or  to  the  influence  of  light,  which  was  entirely  disregarded  in 
all  their  earlier  experiments.  In  addition  Hess  and  Unger  reported, 
that  they  observed  rickets  in  children  fed  on  milk  (600  to  700  cc. 
daily)  containig  2.5  to  3  per  cent  fat,  and  also  in  breast-fed 


RICKETS  327 

children,  whose  mothers  apparently  had  sufficient  vitamine  A  in  the 
diet.  All  of  these  cases  were  cured  with  cod-liver  oil.  Holt, 
Courtney  and  Fales  (1132)  kept  a  child  for  5  weeks  on  vegetable 
fat,  as  the  sole  fat  constituent  (apparently  free  from  vitamine 
A).  The  child  stopped  growing  but  remained  in  good  general 
condition.  Two  children  developed  styes  and  two  others,  facial 
eczema,  which  disappeared  when  milk  fat  was  given,  v.  Groer 
(1133)  conducted  an  experiment  with  two  children,  who  grew  almost 
normally  during  the  first  half  of  the  first  year  on  a  fat-free  diet. 
The  diet  consisted  of  skim-milk,  containing  only  0.01  per  cent  fat, 
and  cane  sugar;  later,  oatmeal  was  added.  Here  we  see  again  the 
demonstration  that  under  certain  conditions  children  may  thrive 
with  very  little  A-vitamine. 

Hess  (I.e.  636),  on  the  other  hand,  could  observe  rickets  in  about 
75  per  cent  of  the  children  on  a  diet  composed  of  "Eiweiss"  milk 
supplemented  with  cream,  dextrin,  malt-soup  and  orange  juice.  He 
admitted,  however,  that  this  diet  was  also  poor  in  vitamine  B. 

Among  the  opponents  of  the  vitamine  theory  of  rickets  (lack  of 
vitamine  A),  we  find  at  first  McCollum,  Simmonds  and  Parsons 
(1134),  on  the  basis  of  their  rat  experiments,  and  also  Robb  (1135). 
The  latter  concluded,  from  guinea  pig  experiments,  that  vitamine  C 
affects  the  mineral  metabolism,  so  that  the  assumption  of  the  existence 
of  a  special  antirachitic  vitamine  does  not  appear  to  be  necessary. 
Later,  McCollum  and  co-workers  (1135a),  Sherman  and  Pappenheimer 
(I.e.  1112b  and  c)  and  Hess,  McCann  and  Pappenheimer  (1135b) 
conducted  some  preliminary  experiments  with  rats  on  diets  deficient 
in  vitamine  A  and  in  other  respects,  and  produced  a  rickets-like 
condition  suggestive  of  a  causative  relationship  of  this  condition  to 
rickets  in  man.  In  1915,  we  noted  in  rats  a  disease  similar  to 
rickets;  a  scoliosis-like  curvature  of  the  spine  with  rosary  develops 
sometimes  in  these  animals  on  diets  very  rich  in  fat  and  containing 
also  cod  liver  oil  (I.e.  331  and  p.  339).  McCollum  and  his  co-workers 
fed  rats  on  artificial  food  mixtures  and  found  that  a  diet  poor  in 
vitamine  A  and  phosphorus,  but  rich  in  calcium,  led  to  rickets. 
A  simultaneous  addition  of  cod  liver  oil  (but  not  butter)  made  the 
diet  satisfactory  for  the  animals.  They  found  that  cod  liver  oil 
specifically  influenced  the  bone  development;  the  deposition  of 
calcium  in  the  bones  could  be  readily  seen  in  the  X-ray  in  from  two 
to  eight  days. 


328  THE   VITAMINES 

Hess,  McCann  and  Pappenheimer  (I.e.  1135b)  maintain  that  a 
lack  of  vitamine  A  is  not  followed  by  rickets  in  rats.  This  is  con- 
trary to  the  findings  of  Mellanby  in  dogs,  and  shows  perhaps  that 
butter  is  very  poor  in  vitamine  A  [in  agreement  with  the  observa- 
tions of  Zilva  and  Miura  (I.e.  574a)]  and  that  dogs  and  rats  show 
great  quantitative  differences  with  respect  to  their  vitamine  A 
requirements  necessary  to  influence  bone  growth.  In  this  con- 
nection, the  vitamine  A  of  butter  is  less  resistant  than  cod  liver  oil 
towards  oxidation.  In  confirmation  of  this,  there  is  the  work  of 
Turk  (1135c)  that  melted  butter  exposed  to  oxidation  does  indeed 
increase  the  immunity  of  children  towards  infections  but  does  not 
protect  against  rickets. 

In  a  recent  publication,  McCollum,  Simmonds,  Shipley  and  Park 
(1135d)  have  attributed  to  cod  liver  oil  a  specific  role  in  rickets. 
The  vitamine  contained  therein  doesn't  seem  to  be  identical  in  its 
influence  with  vitamine  A,  but  perhaps  it  is  a  question  of  quantita- 
tive relationships.  On  the  addition  of  cod  liver  oil  the  calcium  and 
phosphorus  requirements  of  the  animals  were  markedly  reduced. 
A  similar  result  was  obtained  on  the  administration  of  phosphorus. 

On  a  critical  consideration  of  the  vitamine  theory  of  rickets  it  is 
apparent  that  on  a  certain  diet  which  must  be  regarded  as  leading  to 
rickets,  not  all  children  develop  the  disease.  On  the  other  hand,  a 
large  percentage  of  the  cases  may  be  cured  by  cod  liver  oil,  but  not  all 
cases.  Individual  differences  arise  here  which  can  not  be  explained 
at  present,  though  we  firmly  believe  that  we  are  on  the  right  track. 
Recently,  of  the  French  investigators,  Nathan  (1136)  has  adopted 
our  view. 

Undoubtedly,  rickets  may  be  brought  into  causal  relationship  with 
vitamine  A,  although  the  above-mentioned  factors  are  also  of  signifi- 
cance. In  accord  with  this,  there  is  the  fact  that  the  manifestations 
of  rickets  in  children,  in  certain  periods  of  the  year,  are  more  marked 
than  in  others.  Sometimes,  as  Dr.  Hess  has  told  us,  spontaneous 
cures  occur  in  spring.  One  might  be  tempted  to  associate  this  with 
seasonal  variations  in  the  vitamine  content  of  milk,  as  has  already 
been  partially  demonstrated.  The  influence  of  light  is  also  to  be 
considered  in  this  respect.  Hess  and  Unger  believe  in  the  specific 
action  of  cod  liver  oil,  but  not  of  milk  fat.  We  cannot  agree  with 
this  since  it  would  mean  that  normal  milk  contains  no  substance 
protective  against  rickets,  and,  therefore,  most  milk-fed  children 


RICKETS  329 

would  suffer  from  this  disease.  If  we  accept  the  idea,  however, 
that  cod  liver  oil  contains  the  specific  substance,  then  we  admit  that 
the  diet,  which  brings  about  rickets,  must  be  lacking  in  this  sub- 
stance. But  since  butter  and  cod  liver  oil  play  the  same  part  in 
rat  feeding,  we  have  to  recognize  that  rickets  is  caused  by  a  lack 
of  vitamine  A.  We  must  accept  this  logical  relationship,  if  we  wish 
to  proceed  to  the  experimental  tests  of  the  above  conception.  In  the 
meantime,  it  seems  to  us  that  the  study  of  rickets  is  more  likely 
to  lead  to  the  desired  end  if  dogs  are  used  in  preference  to  rats,  for 
in  the  latter  only  osteoporosis  frequently  develops.  The  develop- 
ment of  this  disease  in  young  dogs  shows  an  appreciable  similarity 
to  rickets  in  small  children.  With  the  elucidation  of  the  influence 
of  light,  phosphorus  and  particularly  vitamine  A,  we  may  hope  for  a 
solution  of  the  problem  of  rickets. 

OSTEOMALACIA 

Late  rickets,  which  we  have  already  mentioned,  forms  the  natural 
stepping-stone  between  rickets  in  small  children  and  osteomalacia. 
The  latter  seems  to  be  due  to  an  identical  metabolic  disturbance. 
Some  demonstration  for  the  genetic  relationship  is  provided  by  Ogata 
(1 137) .  In  the  province  of  Toyama  in  Japan,  osteomalacia  developed 
in  adults,  under  conditions  apparently  similar  to  those  giving  rise 
to  rickets  in  children.  Before  the  war,  the  disease  developed  almost 
exclusively  in  young  pregnant  women,  in  which  condition  it  often 
assumed  a  serious  aspect.  This  form  of  the  disease  was  described  by 
Liesegang  (1138)  and  also  by  Scipiades  (1139),  and  was  mostly 
associated  etiologically  with  the  endocrine  glands.  A  relationship 
to  the  pregnancy  was  often  suspected  although  this  condition 
undoubtedly  was  only  a  secondary  factor.  It  is,  however,  not 
impossible  that  an  interruption  of  the  pregnancy  might  be  accom- 
panied by  an  improvement  Koltonski  (l!39a).  Januszewska  (1140) 
reported  on  3500  cases  in  Bosnia  on  an  inadequate  diet,  accompanied 
by  rheumatic  manifestations.  The  cases  appeared  between  Decem- 
ber and  April  and  showed  a  periodicity  similar  to  that  already  noted 
in  rickets,  and  which  was  associated  with  improvement  in  the  diet. 
The  disease  was  favorably  influenced  by  cod  liver  oil. 

During  the  war,  central  Europe  afforded  thousands  of  such  cases. 
This  has  been  observed  in  Vienna,  especially  during  the  summer  of 


330  THE   VITAMINES 

1919  (1141).  The  frequency  of  the  cases  was  remarked  upon  by 
Partsch  (1142)  and  by  Sauer  (1143).  The  disease  did  not  occur  only 
in  the  adolescent  period  (12  to  20  years),  according  to  Hamel  (1144) 
and  Heyer  (1145),  but  also  in  men,  according  to  Kopchen  (1146) 
and  in  old  age,  among  men  and  women,  according  to  Curschmann 
(1147). 

Symptoms 

Chelmonski  (1147a)  describes  two  forms  of  alimentary  bone 
disease  which  prevailed  in  Poland  in  1917-1918;  one  form  was 
accompanied  by  pain  in  the  joints  and  brittleness  of  the  bones; 
the  other,  more  like  the  classical  osteomalacia,  but  in  the  absence 
of  pregnancy,  with  severe  changes  in  the  pelvis  and  progressive 
paralysis  of  the  lower  extremities.  As  in  rickets,  the  bone  changes 
are  most  prominent;  without  this,  the  diagnosis  is  not  easy  and  the 
disease  is  often  confused  with  rheumatism.  In  women,  according  to 
Croftan  (1148),  fractures  appear  chiefly  in  the  pelvic  region.  The 
condition  was  described  by  Looser  (1149)  in  young  people  of  both 
sexes  (17  to  20  years  of  age).  The  symptoms  consist  in  an  enlarge- 
ment of  the  thyroid,  fatigue  on  walking,  pains  in  the  knees  and 
ankles.  Pain  in  the  bones  is  one  of  the  earliest  symptoms.  Alwens 
(1150)  observed  26  cases  among  women  between  19  and  72  years  of 
age,  65  per  cent  of  which  occurred  during  and  after  the  climacteric. 
The  weight  of  the  patients  in  most  cases  was  about  100  pounds; 
15  cases  showed  curvature  of  the  spinal  column,  and  in  10  cases, 
spontaneous  fractures  were  noted.  Bohme  (1151)  emphasized,  in 
particular,  the  complete  analogy  between  rickets  and  osteomalacia. 
Imhof  (1152)  believed  that  osteomalacia  is  frequently  accompanied 
by  psychoses  (dementia  praecox).  Gould  (1153)  believed  in  a 
complete  analogy  between  osteomalacia  and  ,  Recklinghausen's 
disease  and  noted  that  besides  an  affection  of  the  peripheral  nerves 
in  the  latter,  still  other  factors  play  a  role;  in  his  opinion,  the  observed 
bone  lesions  point  to  this.  Cases  of  Recklinghausen's  disease  were 
described  also  by  Weiss  (1153a)  and  Comby  (1153b). 

Anatomy 

Pathological  findings  were  described  by  Hanau  (1154)  and  by  Wild 
(1155).  The  latter  observed  changes  in  the  bones,  similar  to  those 
in  rickets,  namely  uncalcified  osteoid  tissue. 


RICKETS  331 

Metabolism 

Scott  (1156)  studied  the  calcium  metabolism  in  the  puerperal 
type  of  osteomalacia.  The  amount  of  calcium  in  the  urine  was 
increased,  while  during  pregnancy  and  lactation,  the  output  was 
decreased.  The  figures  were  as  follows:  normal  CaCl2  per  liter  of 
urine — 1.146  grams;  in  osteomalacia — 1.36  grams;  iu  pregnancy 
and  lactation — 0.988  gram.  The  calcium  content  of  the  blood  is 
increased  in  osteomalacia;  in  pregnancy,  no  variation  from  the 
normal  figure  was  noted;  normal — 0.45  gram  CaO  per  liter;  osteo- 
malacia— 0.58  gram.  Leo  Zunz  (1157)  conducted  metabolism 
experiments  in  puerperal  osteomalacia.  He  found  a  negative  calcium 
balance,  with  a  definite  improvement  on  castration.  Elfer  and 
Kappel  (1157a)  conducted  metabolism  experiments  with  and  with- 
out the  addition  of  extracts  of  endocrine  glands.  The  calcium 
and  magnesium  balances  were  sometimes  negative,  while  the  phos- 
phorus balance  remained  positive. 

Therapy 

The  cod  liver  oil  therapy  has  not  been  used  very  much,  though 
where  it  has  been  used,  the  reports  were  favorable.  Looser,  Cursch- 
mann,  Kopchen,  Hamel  and  many  others,  have  reported  on  this 
subject.  In  most  cases,  a  change  in  diet  was  instituted,  together 
with  the  therapy. 

Etiology 

Hutchinson  and  Patel  (1157b)  observed  numerous  cases  among 
Mohammedan  women  in  Bombay,  and  attributed  the  cause  of  the 
disease  not  to  the  diet,  early  marriage  or  prolonged  lactation,  but 
to  the  inertia  and  lack  of  air — in  accord  with  the  domestication 
theory  of  rickets.  The  description  of  cases  among  men,  in  middle 
and  old  age,  shows  that  the  disease,  as  such,  has  nothing  to  do  with 
the  puerperium.  The  cases  in  old  age,  as  well  as  in  middle  age,  show 
that  this  rachitic-like  condition  has  nothing  in  common  with  growth, 
contrary  to  the  findings  of  Mellanby  on  rickets  in  young  dogs. 
Recently,  the  prevalence  of  this  disease  in  central  Europe  was 
coincident  with  a  lack  of  milk,  eggs,  cheese  and  butter.  The  popu- 
lation lived  on  cabbage,  turnips,  some  potatoes  and  meat.  There 
was,  therefore,  a  lack  of  vitamine  A,  as  in  rickets. 


CHAPTER  IV 

SOME  NUTRITIONAL  DISTURBANCES  IN  CHILDREN — TETANY,  CARBO- 
HYDRATE DYSTROPHY  (MEHLNAHRSCHADEN),  ATROPHY 

In  the  description  of  these  conditions,  we  must  state,  first  of  all, 
that  under  this  heading,  besides  conditions  that  really  are  avitam- 
inoses,  others  have  been  included  which  possess,  no  doubt,  a  different 
etiology.  Unfortunately,  at  the  present  status  of  our  knowledge, 
this  can  not  be  avoided. 

TETANY  (SPASMOPHILIA) 

Taking  this  disease  as  an  example,  the  clinical  picture  of  spasmo- 
philia  is  not  absolutely  defined,  and  Luzzatti  (1158)  stated  correctly 
that  it  is  difficult  to  differentiate  between  tetany  and  convulsions, 
especially  because  of  the  atypical  course  of  infantile  epilepsy. 
According  to  Stheehman  and  Arntzenius  (1159),  a  poverty  of 
calcium  may  be  followed  by  clinical  consequences  of  various  kinds. 
The  diagnosis  of  spasmophilia  may  be  simplified  by  testing  the 
electrical  irritability,  but  this  test  is  not  absolutely  certain.  It  is 
now  generally  assumed  that  the  tetanic  conditions  in  children  are 
brought  about  by  the  lack  of  calcium.  That  the  calcium  retention 
is  impaired  in  such  cases,  was  demonstrated  by  Cybulski  (1160). 
Theoretically,  however,  it  is  possible  that  the  calcium-impoverish- 
ment of  the  body  may  take  place  in  many  ways.  It  may  be  asso- 
ciated with  a  lack  of  calcium  in  the  diet,  a  condition  which  may  be 
remedied  by  means  of  calcium  therapy.  The  disease  may  be  related 
to  an  affection  of  the  parathyroids,  in  which  event  an  organo-therapy 
would  be  in  order.  Finally,  it  may  be  associated  with  an  impoverish- 
ment of  the  organism  in  calcium  salts,  as  a  result  of  rachitic  condi- 
tions. Pepper  (1161)  correctly  called  attention  to  this  possibility. 
In  practice,  apparently  all  three  causes  may  be  accounted  for. 

Etiology 

The  lack  of  calcium  as  the  sole  cause  of  the  disease  must  be  quite 
rare,  since  most  of  the  available  data  do  not  report  favorably  on 
calcium  therapy.  When  calcium  therapy  is  spoken  of,  as  in  the 

332 


NUTRITIONAL   DISTURBANCES   IN    CHILDREN  333 

cases  of  Rohmer  and  Vonderweidt  (1161a)  and  Scherer  (llGlb), 
this  therapy  was  for  the  most  part  supplemented  by  cod  liver  oil 
and  phosphorus.  Thiemich  (1162)  found  the  calcium  therapy  alone 
without  effect,  while  Petrone  and  Vitale  (1163)  found  the  parathyroid 
therapy  ineffective  in  their  cases,  though  Pincherle  and  Maggesi 
(1163a)  reported  that  their  tetany  cases  showed  pathological 
changes  in  the  endocrine  glands.  The  third  possibility,  the  calcium 
impoverishment  as  result  of  rachitic  metabolic  disease,  is  strengthened 
by  the  frequent  association  of  this  disease  with  rickets  and  osteo- 
malacia.  Thus,  Januszewska  (I.e.  1140)  found  338  cases  of  tetany 
among  3500  cases  of  osteomalacia.  Sauer  (I.e.  1143),  too,  described 
this  relationship  and  was  strengthened  in  his  view  by  a  favorable 
cod  liver  oil  therapy.  Takasu  (1164)  associated  spasmophilia  with 
beriberi.  In  his  cases,  there  was  a  possibility  of  infantile  beri- 
beri. Since  the  rachitic  disturbance  may  occur  at  any  age,  the 
related  tetany  may  appear  similarly.  Klose  (1165)  believed  that 
this  disease  can  not  occur  under  the  age  of  2  months.  Bliihdorn 
(1166)  on  the  contrary,  found  it  in  children  of  all  ages.  Apart  from 
this,  it  is  not  totally  excluded  that  certain  forms  of  tetany  of  preg- 
nancy and  lactation  are  to  be  attributed  to  the  above  causes,  although, 
this  was  denied  by  Faas  (1167).  Tetany  seems  to  be  favorably 
affected  by  a  one-sided  flour  diet,  and  occurs  rarely  in  breast-fed 
infants,  according  to  Guthrie  (1168).  Lust  (1169)  stated  that  tetany 
occurs  only  in  2  per  cent  of  breast-fed  and  in  55.7  per  cent  of  artifi- 
cially-fed children.  According  to  Bossert  and  Gralka  (1169a)  true 
spasmophilia  may  be  recognized  by  the  fact  that  it  can  be  influenced 
by  a  change  in  the  diet.  v.  Meysenbug  (1170)  did  not  believe  that 
spasmophilia  had  anything  to  do  with  the  three  known  vitamines, 
still  we  cannot  sse  that  his  results  justify  such  far-reaching  conclusions. 
Jeppson  and  af  Klercker  (1170a)  believe  that  tetany  is  not  due  to 
the  lack  of  calcium  in  the  diet  but  to  an  excess  of  phosphorus. 

Symptoms 

The  Erb  sign  (increase  of  galvanic  irritability  of  the  muscles  and 
the  motor  nerves)  and  the  Chvostek  sign  [increase  of  the  irritability 
of  the  nerves  especially  the  facial  nerve  or  the  popliteus  (Ibrahim, 
1170b)  to  mechanical  stimulus]  are  of  value  in  the  diagnosis  of 
tetany.  According  to  Stheehman  and  Arntzenius  (I.e.  1159),  the 
Chvostek  sign  is  more  sensitive.  Bossert  (1171)  occasionally  ob- 


334  THE    VITAMINES 

served  edema  and  carpopedal  spasms.  The  determination  of  the 
calcium  content  of  the  blood  is  of  help  in  the  diagnosis.  Howland 
and  Marriott  (1172)  found,  in  normal  cases,  10  to  11  mgm.  calcium 
in  the  blood;  in  rickets,  somewhat  less;  in  tetany  an  average  of 
5.6  mgm.  and  sometimes  only  3.5  mgm.  They  found  that  the  calcium 
therapy,  even  though  gradual,  is  effective.  Elias  and  Spiegel  (1172a) 
found  the  phosphorus  content  of  the  blood  very  high,  while  Tisdall, 
Kramer  and  Howland  (1172b)  found  that  the  ratio  of  sodium  and 
potassium  to  calcium  and  magnesium  is  very  greatly  increased. 
Stheehman  and  Arntzenius  (1173)  found,  on  the  other  hand,  that 
the  calcium  content  of  the  blood  is  no  definite  indication  of  tetany. 
The  calcium  in  the  blood  may  be  high  and  yet  the  Erb  sign  is  posi- 
tive, which  may  be  associated  with  a  lack  of  calcium  in  the  tissues. 
v.  Meysenbug  (1173a)  states  that  the  Thiemich  sign  is  not  associated 
with  the  low  calcium  content  of  the  blood  and  that  larger  doses  of 
calcium  in  older  children  do  not  affect  the  anodic  reaction.  The 
determination  of  salt  metabolism  in  tetany  was  made  by  Fletcher 
(1174).  Liefmann  (1175)  found  the  urinary  acetone  elimination 
very  high  in  the  acute  stage,  while  Sharpe  (1176)  found  an  increased 
guanidine  elimination  in  the  feces.  Hoobler  (1177)  stated  that  the 
Ca-  Mg-  and  P  balances  were  all  negative.  Bolten  (1177a)  found 
a  decreased  blood  calcium  content  with  an  increase  in  the  electrical 
sensitivity  in  Fragilitas  ossium  (Osteopsathyrosis) . 

Therapy 

Calcium  therapy  was  described  by  Howland  and  Marriott  (I.e. 
1101).  Thiemich  (I.e.  1162)  recommended  breast  feeding  and 
phospho-cod  liver  oil.  Brown  and  Fletcher  (1178)  also  recom- 
mended phospho-cod  liver  oil.  Stheehman  (1179)  and  Brown, 
MacLachlan  and  Simpson  (1180)  have  corroborated  the  cod  liver 
oil  therapy.  The  last  named  workers  found  the  intravenous  injec- 
tion of  calcium  lactate  without  effect  if  not  accompanied  by  a  simul- 
taneous phospho-cod  liver  oil  therapy.  The  clinical  manifestations 
of  tetany  disappear  in  10  to  17  days  while,  without  the  addition  of 
calcium,  the  action  is  very  much  delayed.  Unfortunately,  we  have 
found  no  data  on  cod  liver  oil  therapy  alone.  Recently,  Huldschin- 
sky  (1180a)  has  used  in  tetany  the  sun-ray  therapy  with  which  he 
was  successful  in  the  treatment  of  rickets.  It  appears  that  the  sub- 
ject requires  still  further  experimental  study. 


NUTRITIONAL   DISTURBANCES   IN   CHILDREN  335 

CARBOHYDRATE   DYSTROPHY    (MEHLNAHRSCHADEN) 

It  is  evident  that  under  this  name  conditions  are  classified,  the 
etiology  of  which  is  fundamentally  different.  In  1905-1910,  par- 
ticularly in  Germany,  but  also  in  other  countries,  widespread  diseases 
of  children  appeared.  These  arose  as  the  result  of  proprietary  foods 
fed  over  a  long  period  of  time.  Czerny  (1181)  was  first  to  recognize 
the  dangers  of  this  regime,  and  called  the  resultant  conditionr 
"Mehlnahrschaden."  It  need  hardly  be  said  that  the  disease  occurred 
only  in  artificially  fed  babies.  Children  who  could  tolerate  cow's 
milk  poorly  or  not  at  all,  were  fed  for  weeks,  more  often  for  months, 
on  various  proprietary  foods.  Recently,  Bloch  (1181a)  also  warned 
against  the  danger  of  a  carbohydrate-rich  diet  in  infants.  At  first, 
this  food  seems  to  be  well  tolerated;  the  splendid  appearance  of  the 
child  permits  of  no  suspicion  that  the  food  in  inadequate.  Later, 
however,  after  a  lapse  of  time  which  varies  within  wide  limits  in 
different  children,  a  disease  develops  which,  according  to  Rietschel 
(1182),  occurs  in  three  different  forms:  pure  atrophic  (hunger), 
atrophic-hydremic  and  hypertonic. 

The  pure  atrophic  form  occurs  in  children  fed  on  flour,  with  the 
following  symptoms.  The  stools  react  acid,  with  considerable  slimy 
substance  and  fermentation.  At  the  same  time,  there  develops  a 
severe  atrophic  hypertonicity  of  the  musculature;  the  muscles  are 
tightly  stretched,  which  simulates  a  healthy  condition.  Electrical 
supersensitivity  of  the  peripheral  nerves  is  frequently  noted.  Here, 
there  is  often  a  great  susceptibility  to  secondary  infections  (thrush, 
aphthae,  abscesses  of  the  skin,  otitis;  phlegmons,  and  pneumonia). 
In  spite  of  the  severe  symptoms,  the  gastro-intestinal  tract,  on 
autopsy,  is  found  almost  intact. 

The  atrophic-hydremic  form  manifests  itself,  at  the  beginning,  in 
edema  of  the  lower  extremeties,  which,  according  to  Noeggerath 
(1183)  is  due  to  heart  weakness;  later,  the  children  show  a  blown-up 
face,  sometimes  also  scorbutic  changes  of  the  gums  (Bogen,  1184). 

The  hypertonic  form  resembles  peripheral  nerve  affections  very 
much.  In  this  category,  we  find  cases  where  the  legs  are  drawn  up 
to  the  abdomen,  with  rigid  spinal  column,  tetany  with  increased 
electrical  irritability  and  carpopedal  spasms  lasting  for  weeks.  The 
prognosis  is  bad  and  the  children  die  with  neuritis  (Grimeberg,  1185). 


336  THE   VITAMINES 

Etiology 

Langstein  (1186)  believed  in  the  analogy  of  this  disease  with 
beriberi.  In  one  case,  he  saw  such  a  great  hypertonus  of  the  muscles 
that  he  thought  it  might  be  a  case  of  Little's  disease.  Some  supposed 
cases  of  carbohydrate  dystrophy  may  really  have  been  beriberi. 
According  to  Hess  (I.e.  918),  who  noted  the  development  of  the 
disease  in  the  presence  of  sufficient  vitamine  C,  an  analogy  with 
scurvy  is  excluded.  On  the  other  hand,  according  to  some  recent 
work,  it  seems  to  be  similar  to  hunger  edema.  L.  F.  Meyer  (1187) 
and  Rietschel  (1188)  believed  this  to  be  true.  Following  this, 
Benjamin  (1189)  reported  that  in  carbohydrate  dystrophy,  after 
feeding  "Eiweiss"  milk  there  was  high  nitrogen  retention,  pointing 
perhaps  to  a  craving  of  the  organism  for  protein.  At  first,  the  reten- 
tion amounted  to  50  per  cent  of  the  intake;  later,  15  to  20  per  cent. 
Since  the  cause  of  hunger  edema  has  often  been  regarded  as  lack  of 
protein,  this  statement  is  of  interest.  However,  since  a  diet  com- 
posed chiefly  of  flour  may  be  very  poor  in  vitamines,  under  certain 
conditions,  the  possibility  of  dealing  with  a  vitamine  deficiency  can 
not  be  excluded.  Hess  (I.e.  918)  also  came  to  this  conclusion.  At 
any  rate,  Keller  (1190)  is  right  when  he  says  that  an  almost  exclusive 
carbohydrate  diet  may  be  used  only  intermittently.  However,  there 
seem  to  be  cases  in  the  literature,  as  that  of  Steinitz  and  Weigert 
(1191),  in  which,  as  a  result  of  a  poor  tolerance  for  milk,  an  exclusive 
carbohydrate  diet  was  given  for  a  long  time.  Salge  (1192)  believed 
that  in  individual  children,  there  are  great  differences  in  the  tolerance 
for  starch.  He  saw  a  child  fed  for  2  months  on  flour  exclusively 
without  any  untoward  results.  The  pathological  conditions  arising 
from  a  diet  rich  in  starch  were  studied  also  by  Abt  (1193). 

ATROPHY 

In  the  English  pediatric  literature  the  term  "Mehlnahrschaden"  is 
rarely  encountered;  disturbances  of  this  kind  are  generally  classified 
as  "atrophy"  (Cautley,  1194).  The  latter  described  the  symptoms 
as  follows:  The  disease  arises  as  a  result  of  an  inadequate  diet, 
especially  one  abundant  in  flour.  There  is  considerable  loss  of 
weight,  diarrhea  and  vomiting.  These  symptoms  are  frequently 
accompanied  by  stomatitis  aphthosa,  enteritis,  bronchitis  and  furun- 
culosis.  In  severe  cases,  he  found  edema  of  the  skin  of  the  extrem- 


NUTRITIONAL   DISTURBANCES   IN   CHILDREN  337 

ities,  later  of  the  face,  petechiae  and  opisthotonos.  On  post-mortem, 
edema  of  the  brain  membranes,  pleural  ecchymoses,  erosions  in  the 
stomach  and  intestine,  and  subepithelial  hemorrhages  were  found. 
In  chronic  cases,  there  was  fatty  degeneration  of  the  liver  and  kidney 
epithelium. 

Frank  and  Stolte  (1195)  analyzed  the  organs  of  children  who  had 
died  of  carbohydrate  dystrophy,  special  attention  being  given  to 
the  liver.  This  was  found  to  be  higher  in  sodium  and  chlorine  and 
lower  in  potassium,  while  fat,  ash,  magnesium,  calcium,  phosphorus 
and.  sulphur  showed  no  great  deviations  from  the  normal.  Steinitz 
and  Weigert  (I.e.  1191)  noticed,  in  one  case,  a  high  fat  content  of  the 
organs,  and  more  P205. 

Noeggerath  (I.e.  1183)  and  Hohlfeld  (1196)  recommended  the 
administration  of  raw  milk.  Bendix  (1197)  recommended  breast 
feeding  for  atrophy. 

OTHER  NUTRITIVE   DISTURBANCES   IN   CHILDREN 

In  many  well  known  nutritional  disorders  of  infants,  the  present 
pediatric  literature  gives  no  clear  picture  of  the  etiology.  Above 
all,  the  individual  pictures  are  not  sharply  differentiated.  Morse 
and  Talbot  (1198)  have  remarked  upon  the  chaotic  condition  in  the 
present  conception  of  atrophy.  It  is  therefore  all  the  more  difficult 
for  one  unacquainted  with  this  subject  to  have  a  definite  conception 
as  regards  the  nature  of  this  condition.  Of  the  nutritional  disorders 
to  be  treated  here,  there  is  exudative  diathesis,  atrophy  (in  the  sense 
of  the  German  pediatric  literature),  marasmus,  milk  dystrophy,  etc. 
Many  of  these  conditions  are  now  thought  to  be  due  to  excess  of  fat. 
In  children  practice,  cases  are  known  in  which  every  possible  dietary 
composition  has  been  tested,  but  without  result;  there  are  even 
children  (though  seldom)  who  can  no  longer  tolerate  a  completely 
normal  breast  diet.  How  far  this  may  be  due  to  organic  disturb- 
ances, or  to  a  previous  unsuitable  diet,  is  not  always  certain.  In 
some  cases,  the  deficient  diet  might  have  caused  such  great  harm, 
that  the  child  can  no  longer  be  helped,  even  by  the  best  of  diets. 
Marriott  (1198a)  differentiated  two  forms  of  nutritive  disturbances 
in  infants,  one  form  arising  after  severe  diarrhoea  and  another 
manifested  as  chronic  malnutrition  (marasmus). 

For  some  of  the  above  mentioned  nutritional  disturbances,  Pehu 
(1199)  introduced,  in  France,  vegetable  soups  as  a  therapeutic 


338  THE   VITAMINES 

measure.  This  therapy  was  used  in  Germany  by  Moro  (1200),  in 
the  form  of  carrot  soup,  with  good  results.  A  similar  therapy  was 
recommended  by  Stark  (1201)  in  suspected  avitaminoses,  and 
recently,  there  has  been  a  similar  report  by  Aron  and  Samelson  (1202). 
McClendon  and  Sedgwick  (1202a)  used  spinach  powder  in  infant 
nutrition  and  obtained  satisfactory  increases  in  weight.  Aron 
(1202b)  made  use  of  the  soluble  flour  obtained  by  the  action  of 
ferments  and  found  that  such  products  were  well  utilized  in  the 
more  severe  cases. 

From  time  to  time,  there  appear  in  the  literature  statements  as  to 
a  successful  vitamine  therapy  in  certain  disturbances.  Kohlbrugge 
(1203)  successfully  treated  cholera  infantum,  a  disease  generally 
regarded  as  an  infection  or  intoxication,  with  orange  juice.  Holt, 
Courtney  and  Fales  (1204)  treated  a  case  of  intestinal  infantilismus 
with  cod  liver  oil,  with  good  results. 

It  need  hardly  be  said  that  in  the  feeding  of  children,  the  same 
rules  are  to  be  observed  as  have  already  been  formulated  for  animals. 
Besides  the  already  known  dietary  constituents,  we  must  provide  for 
the  presence  of  the  three  known  vitamines.  It  is  also  essential  that 
the  more  important  constituents  should  not  occur  in  the  diet  in  too 
great  a  dilution.  Some  of  the  recorded  therapeutic  results  with 
vitamines  depend  upon  the  fact  that  either  the  certain  vitamines, 
although  they  were  present,  were  not  sufficient,  or  the  relative  propor- 
tion of  the  individual  food  constituents  to  each  other  was  not  quite 
correctly  chosen.  Weill  and  Mouriquand  (1205)  have  pointed  out 
such  a  possibility;  as  regards  vitamine  B  in  this  connection,  we  have 
the  work  of  Daniels,  Byfield  and  Loughlin  (1206).  We  may  explain, 
in  this  way,  certain  results  of  vitamine  therapy  in  an  insufficiently 
defined  condition,  usually  called  "marasmus"  or  "malnutrition." 
Eddy  and  Roper  (1207),  and  Dubin  and  Lewi  (I.e.  628)  obtained 
marked  results  in  the  conditions  just  mentioned. 

As  regards  atrophy,  it  is  caused  by  excess  of  fat,  according  to  the 
present  conception.  The  results  of  Friedberg  and  Noeggerath  (1208) 
are  to  be  regarded  in  this  light;  they  recommended  the  use  of  centri- 
fuged  breast  milk  in  this  condition.  This  recalls  certain  investiga- 
tions on  rats,  made  by  Funk  and  Dubin  (I.e.  331).  These  animals 
received  a  complete  synthetic  diet,  containing  more  than  50  per 
cent  fat.  The  young  rats  were  able  to  live  on  this  diet  for  2  to  3 
months.  In  spite  of  the  vitamine  content  of  the  diet,  the  animals 


NUTRITIONAL   DISTURBANCES   IN    CHILDREN  339 

did  not  grow;  they  lost  about  50  per  cent  of  their  weight  and  their 
condition  was  such  as  is  mostly  described  in  atrophic  children.  The 
animals  showed  a  fatty  degeneration  of  the  liver;  in  one  case,  there 
was  edema  and  scoliosis,  with  a  permanent  curvature  of  the  spine. 
A  physiological  explanation  of  this  appearance  is  still  to  be  found. 
Should  it  appear,  however,  that  the  milk  dystrophy  and  the  decom- 
position of  Finkelstein  are  attributable  to  the  high  fat  content  of  the 
diet,  then  the  influence  of  fat  may  be  studied  experimentally  in  the 
above  manner. 

In  infantile  atrophy,  Salge  (1209)  found  the  assimilative  capacity 
satisfactory.  The  energy  and  metabolism  of  these  children  was 
studied  by  Bahrdt  and  Edelstein  (1210).  The  assimilation  of  fat 
was  found  normal  by  Hutchinson  (I.e.  1096) ;  fat  losses  are,  however, 
quite  high  as  a  result  of  voluminous  stools.  The  protein  content  of 
the  blood  was  found  low  by  Utheim  (1211)  and  he  believed  that  the 
protein  synthesis  proceeds  abnormally;  in  other  cases,  on  the  con- 
trary, there  may  have  been  an  excessive  carbohydrate  nutrition. 
Matt  ill,  Mayer  and  Sauer  (1212)  state  that  the  glucose  tolerance  of 
atrophic  children  is  higher  than  normally.  Gladstone  (1213)  recom- 
mended a  fruit-juice  therapy.  Cure  with  breast  feeding,  and  in 
one  case  with  cow's  milk,  was  described  by  Marfan  (1213a). 

The  exudative  diathesis  of  Czerny  (1214)  is  not  associated  with 
lack  of  vitamines,  according  to  Hess  (I.e.  918).  The  condition, 
however,  leads  frequently  to  scurvy,  eczema  and  petechial  hemor- 
rhages. According  to  Czerny,  this  disease  arises  because  of  the 
harmful  effect  of  the  dietary  fat.  Schippers  (1215)  found,  in  these 
conditions,  a  low  fat  content  in  the  blood.  Marfan  (1215a)  regards 
athrepsia  as  a  deficiency  disease  but  not  as  an  avitaminosis,  and  he 
accepts  as  the  cause  a  lack  of  specific  enzymes  in  breast  milk.  Be- 
cause of  the  present  better  understanding  of  the  nutritive  require- 
ments of  infants,  a  fundamental  repetition  of  these  nutritional 
problems  is  greatly  to  be  desired. 


CHAPTER  V 

NUTRITION  IN  MAN — AN  INTRODUCTION  TO  THE  STUDY  OF  PELLAGRA 
AND  HUNGER  EDEMA 

From  what  has  been  brought  out  till  now,  it  is  clear  that  in  nutri- 
tion, it  is  not  sufficient  that  all  the  known  dietary  constituents  be 
present;  they  must  also  bear  the  proper  relationship  to  each  other. 
This  idea  developed  chiefly  through  observations  on  rats.  If  these 
animals  are  given  a  qualitatively  correct  diet  (ad  libitum),  dietary 
deficiencies  develop  nevertheless.  It  would  be  tempting  to  assume 
that  these  animals  need  only  ingest  more  food  in  order  to  receive 
all  of  the  constituents  in  correct  amounts.  In  practice,  however,  it 
is  evidently  not  the  case.  No  matter  how  much  food  the  animal  eats, 
the  relationship  of  the  individual  components  is  not  altered.  This 
excessive  dilution  of  certain  components  may  appear  in  human  nutri- 
tion, and  yield  a  rather  unclear  etiological  picture. 

Pellagra,  sprue,  hunger  edema,  and  perhaps  carbohydrate  dys- 
trophy in  children,  may  be  associated  with  the  above  condition.  At 
least  two  of  the  diseases  mentioned  have  recently  been  attributed  to 
the  lack  of  a  protein  of  high  biological  value.  A  number  of  animal 
proteins  are  generally  regarded  as  such.  Should  this  conception 
prove  to  be  correct,  then  these  pathological  conditions  must  be 
stricken  from  the  list  of  avitaminoses,  which  has  in  fact  already 
been  done  by  some  investigators.  We  feel,  however,  that  these 
diseases  have  been  cleared  up  so  little,  etiologically,  that  this  pro- 
cedure seems  to  us  altogether  unjustified  at  present.  In  the  last 
war,  nutrition  played  a  greater  role  than  has  been  generally  assumed 
by  the  laity.  At  the  time  of  the  war,  we  saw  one  immense  metabolic 
experiment,  from  which  a  great  number  of  conclusions  could  have 
been  drawn,  if  the  serious  situation  had  not  prohibited  a  cold-blooded 
survey.  The  late  pharmacologist,  Meltzer,  remarked  at  one  time 
that  all  scientific  data  obtained  during  the  war  would  have  to  be 
corroborated  after  the  war.  He  may  have  been  correct  in  this  state- 
ment, though  the  convincing  power  of  the  large  involuntary  nutri- 
tional experiment  remains  unquestioned,  and  we  may  draw  a  number 
of  conclusions  for  our  purpose.  Various  problems  must  be  taken 
into  account  in  this  connection,  namely: 

340 


NUTRITION   IN   MAN  341 

1.  The  question  of  the  protein  minimum. 

2.  Comparison  of  the  nutritive  value  of  vegetable  and  animal 
protein. 

3.  The  chemical  groups  in  protein  responsible  for  its  high  biological 
value. 

4.  Influence  of  war  food  on  the  health  of  the  people. 
Unfortunately,  because  of  lack  of  space,  we  can  not  enter  into  a 

detailed  discussion  of  the  above  questions.  Protein  has  always  been 
given  an  exceptional  place  in  the  system  of  nutrition  and  correctly 
so.  Recently,  this  has  bee/n  brought  out  in  the  work  of  Gigon  (1216) 
and  Grumme  (1217) .  It  is  quite  important  to  know  the  physiological 
protein  minimum,  and  in  this  respect,  there  are  the  views  held  by 
two  schools — that  of  Voit-Rubner,  which  requires  a  high  protein 
minimum,  and  that  of  Chittenden-Hindhede,  which  is  satisfied  with 
a  considerably  smaller  figure.  The  figures  of  Chittenden  were 
appreciably  reduced  by  Hindhede  (l.o*  732),  who  showed  (1218)  in  a 
great  number  of  extended  nutrition  experiments,  that  adults  could 
maintain  themselves  and  work  normally  on  a  daily  quota  of  20  grams 
of  assimilated  protein  and  3900  calories.  The  investigations  were 
continued  for  a  longer  time  than  usual  for  experimental  purposes 
and  therefore  are  very  convincing.  In  one  instance,  the  experiment 
lasted  305  days  and  the  diet  consisted  of  potatoes  (cf.  p.  255),  mar- 
garine, and  sometimes  onions  and  fruits.  Hindhede  believed  that 
when  the  diet  contains  sufficient  calories,  one  does  not  need  to  worry 
about  the  protein  content,  taking  care  only  that  the  diet  should 
contain  sufficient  vitamines.  The  bread  experiments  of  Hindhede 
(1219)  fall  in  the  same  category.  Of  the  bread,  made  from  whole 
wheat,  30  to  35  grams  assimilated  protein  were  necessary.  The  rest 
of  the  diet  consisted  of  margarine  and  fruits,  and  the  longest  experi- 
ment lasted  242  days.  One  experiment  with  coarsely  smashed  wheat 
(I.e.  791)  lasted  282  days.  In  some  of  these  investigations  it  is  even 
noteworthy  that  the  persons  submitting  to  the  experiment  did  not 
develop  scurvy,  though  it  is  possible  that  fruits  were  used.  The 
minimum  protein  figure  was  very  low  in  experiments  with  barley- 
water  gruel  (1220),  with  the  addition  of  margarine  and  sugar. 
Hindhede  (1221)  also  studied  the  fat  minimum  in  nutrition  which  is 
likewise  of  importance  in  the  determination  of  the  value  of  war  diets. 
In  this  case,  the  experimental  diet  was  composed  of  potatoes,  cabbage, 
rhubarb  and  apples,  but  without  addition  of  fat.  The  experimental 


342  THE   VITAMINES 

subjects  were  in  splendid  health  for  16  months,  so  that  a  lack  of 
animal  protein  and  fat  can  not  be  regarded  as  the  cause  of  pellagra 
and  war  edema,  since  they  were  on  this  diet  much  longer  than  is 
required  for  the  development  of  this  disease.  Bang  (1222)  believes, 
however,  that  Hindhede's  experimental  subjects  are  exceptions  and 
hence  his  results  can  not  be  used  without  further  confirmation, 
especially  as  regards  children  and  consumptives. 

Hindhede  assumed  that  a  gram  of  potato  or  bread  protein  when 
assimilated,  corresponds  exactly  to  a  gram  of  meat  or  milk  protein, 
but  this  conception  has  failed  to  gain  recognition  up  to  the  present. 
In  a  new  series  of  rat  experiments,  McCollum,  Simmonds  and 
Parsons  (1222a)  observed  that  the  internal  organs,  especially  the 
kidney,  can  supplement  effectively  the  diet  of  plant  origin.  Neu- 
mann (1223)  believes  that  it  is  possible  to  live  on  an  exclusive  bread 
diet.  Dollner  (1224),  however,  in  the  description  of  the  etiology  of 
hunger  edema,  called  it  "potato  sickness."  He  saw  this  condition 
develop  on  large  amounts  of  potatoes  and  cereals.  Kraszewski 
(1225)  believes  that  potatoes  are  poor  in  calcium  and  magnesium 
and  hence  must  be  supplemented  with  bread  and  vegetables.  Un- 
fortunately the  individual  investigators  are  not  agreed  as  to  the 
protein  minimum.  Jansen  (1226)  figured  that  for  a  man  weighing 
62  kilos,  2000  calories  and  60.5  grams  protein  are  necessary;  but  the 
diets  used  by  Hindhede  were  much  richer  in  calories.  Sherman 
(1227)  calculated  that  for  an  adult  weighing  70  kilos,  44.4  grams 
protein  per  day  are  necessary.  Taylor  (1228),  in  his  studies  on  the 
nutrition  of  English  prisoners  in  Germany,  concluded  that  70  to  90 
grams  protein  must  be  fed.  Benoit  (1229)  investigated  in  Germany, 
the  metabolism  of  a  number  of  prisoners  (Russian  officers)  and  found 
that  they  maintained  themselves  without  complaint  for  16  months 
on  48  grams  protein  and  1700  calories,  the  protein  being  partly 
vegetable  and  partly  animal.  Botazzi  (1230)  was  opposed  to  the 
introduction  into  the  Italian  army,  of  Chittenden's  figures.  Kruse 
and  Hintze  (1231)  accepted  three-quarters  of  Voit's  food  standards. 
Praussnitz  (1232)  also  discussed  this  matter.  Stille  (1233)  supported 
the  views  of  Hindhede  against  those  of  Rubner.  Hindhede  recorded 
the  diet  on  which  he  and  his  family  lived  during  the  war,  which 
consisted  of  54.6  grams  protein  and  2100  calories.  Funk,  Lyle  and 
McCaskey  (1234)  reported  a  20  day  metabolism  experiment  on  four 
men,  receiving  yeast,  rice  and  white  bread.  A  nitrogen  balance  was 


NUTRITION   IN   MAN  343 

almost  obtained  with  5  to  7  grams  nitrogen.  The  somewhat  unsatis- 
factory results  were  perhaps  due  to  a  partial  absence  of  vitamines 
(especially  during  the  periods  without  yeast),  or  to  a  relatively  low 
caloric  content.  This  was  one  of  the  few  metabolism  experiments 
where  the  effect  of  such  a  dietary  mixture  on  the  composition  of  the 
blood  was  studied.  As  to  the  nutritive  value  of  bakers'  yeast, 
Hawk,  Smith  and  Holder  (1235)  reported  favorably  that  yeast  can 
replace  30  per  cent  of  the  protein  of  the  diet. 

After  this  brief  consideration  of  the  protein  minimum,  we  come  to 
the  comparison  between  the  nutritive  value  of  vegetable  and  animal 
protein.  That  there  is  a  great  difference  in  favor  of  the  animal 
protein,  is  now  maintained  by  many,  in  opposition  to  the  view  held 
by  Hindhede.  The  present  view  of  Hindhede  was  advocated  many 
years  ago  by  Rutgers  (1236).  Boruttau  (1237)  showed  in  metabo- 
lism experiments,  that  vegetable  proteins  are  well  utilized;  he  did 
not  believe  that  the  biological  value  was  an  indication  of  the  vitamine 
content,  for  unpolished  rice  has  a  lower  biological  value  than  polished 
rice.  Botazzi  (1238),  on  the  contrary,  believed  in  the  special  nutri- 
tive value  of  animal  protein  and  fat.  Rose  and  Berg  (1239)  proposed 
a  view,  approaching  ours,  in  that  they  assumed  that  the  protein 
requirements  depend  on  the  nature  of  protein  and  the  composition 
of  the  rest  of  the  diet.  Milk  protein  has  mostly  been  given  an 
exceptional  place  in  nutrition.  Recently,  this  was  again  reported 
upon  by  Hart  and  Steenbock  (1240),  and  we  must,  therefore,  ask 
ourselves  the  question,  which  chemical  group  in  the  protein  molecule 
plays  such  an  extraordinary  role  in  nutrition. 

This  problem  has  been  treated  experimentally  in  such  great  detail, 
that  a  book  may  well  be  written  on  the  subject.  For  this  reason, 
we  must  confine  ourselves  to  the  chief  results.  The  biological  value 
of  proteins  has  been  determined  by  Thomas  (I.e.  1)  on  different 
products  and  yielded  the  following  figures : 

Beef 104      Rice 88      Peas 56 

Cow'smilk 100      Potatoes 79      Wheat  flour 40 

Fish 95      Casein 70      Corn  flour 30 

This  table  is  based  upon  the  conception  of  the  similarity  of  animal 
proteins  with  body  proteins  and  is  due  to  their  relative  content  of 
essential  amino  acids.  A  number  of  amino  acids  have  been  recog- 
nized as  such.  Still,  in  the  study  of  the  nutritive  requirements  of 
the  lower  organisms  (yeast)  it  has  been  established  that  some  proteins 


344  THE   VITAMINES 

contain  substances  which,  after  hydrolysis,  cannot  be  isolated 
and  which  thus  far  remain  unknown  among  the  cleavage  products. 
McCollum  and  Davis  (I.e.  97)  believed  that  when  casein  is 
autoclaved,  its  nutritive  value  decreases  markedly;  but,  according  to 
Funk  and  Macallum  (I.e.  98),  this  deficiency  may  be  overcome  by 
the  addition  of  vitamine-containing  products.  Yet,  it  is  not  impos- 
sible that  heating  may  decrease  the  nutritive  value  of  some  proteins. 
Mueller  (I.e.  167a),  in  this  connection,  made  an  interesting  observa- 
tion in  the  culture  of  streptococci,  namely,  that  peptone,  casein, 
edestin,  in  the  tryptophane  fraction,  contains  a  substance  seemingly 
indispensable  for  the  growth  of  this  microorganism.  Mueller  was  of 
the  opinion,  that  he  was  dealing  with  a  new  amino  acid,  in  which 
belief  he  was  strengthened  by  the  fact  that  reprecipitated  casein 
possessed  the  same  ability.  Still,  this  condition  does  not  militate 
against  the  possibility  that  the  substance  may  be  of  a  vitamine  nature, 
for  in  the  first  place,  the  active  substance  is  so  markedly  adsorbable 
that  it  may  be  completely  removed  with  animal  charcoal;  second,  if 
the  substance  is  carried  down  in  the  first  precipitation  of  the  casein, 
it  would,  in  all  probability,  be  carried  down  on  reprecipitation.  By 
reprecipitation  of  the  casein,  therefore,  no  separation  can  be  obtained. 
We  saw  in  the  case  of  other  bacteria  that  the  vitamine-like  substances 
there  could  not  be  filtered,  even  through  paper,  without  being 
adsorbed.  Mueller  found  that  wool,  silk,  gelatin  and  hydrolyzed 
yeast  do  not  contain  this  substance ;  it  was  found  in  the  tryptophane 
fraction;  but  when  the  known  substances  in  this  fraction  were 
tested  for  the  above  action,  no  result  was  obtained.  If  the  fraction 
is  precipitated  with  phosphotungstic  acid,  the  substance  disap- 
pears from  the  precipitate  as  well  as  from  the  filtrate.  If  the 
fraction  is  crystallized,  the  substance  remains  in  the  filtrate  and 
appears  unstable,  de  Souza  and  McCollum  (I.e.  529)  observed 
that  hydrolyzed  meat  exerts  some  action  on  the  growth  of  yeast. 
Funk  and  Dubin .  (unpublished  work)  noted  that  a  completely 
extracted  meat  (the  last  extract  being  no  longer  active  for  yeast), 
when  hydrolyzed,  shows  definite  action.  The  question  whether  we 
are  dealing  here  with  a  vitamine  or  an  unknown  amino  acid  must  be 
left  open  for  the  time  being,  though  it  seems  to  us  not  impossible 
that  the  above  finding  will  give  a  new  stimulus  to  protein  research, 
in  that  we  have  a  biological  reaction  that  may  be  of  value  in  a  frac- 
tionation.  In  addition,  the  nutritive  value  of  the  proteins  may 
appear  to  us  in  a  new  light,  for  the  experiments  show  that  some 


NUTRITION   IN  MAN 


345 


proteins,  in  a  more  or  less  impure  condition  (for  the  degree  of  purity 
can  naturally  not  be  accurately  determined),  besides  the  known 
cleavage  products,  still  contain  something  that  has  been  unknown 
to  us.  It  is  very  tempting  to  associate  this  substance  with  the 
etiology  of  pellagra  and  similar  conditions,  though  we  must  admit, 
that  the  basis  for  it  is  at  present  hardly  at  hand.  This  assumption, 
however,  explains  some  of  the  established  facts  about  pellagra  very 
well,  as  we  shall  presently  see. 


DIET* 

MEAT 

SUGAR 

STARCH 

LARD 

SALTS 

AUTO- 
LTZED 

TEAST 

ORANGE 

A  GAR 

COD 
LIVER 
OIL 

Meat  

grams 

49 

grams 
12 

grams 

12 

grams 
12 

grams 

3 

CC. 

4 

CC. 

3 

grams 

3 

CC. 

5 

Sugar 

12 

49 

12 

12 

3 

4 

3 

3 

5 

Starch 

12 

12 

49 

12 

3 

4 

3 

3 

5 

Lard  

12 

12 

12 

49 

3 

4 

3 

3 

5 

DIET 

WEIGHT—  FIRST 
25  DAYS 

WEIGHT  —  FOLLOW- 
ING 55  DAYS 

TOTAL 
WEIGHT 

INCREASE! 

Increase 

Decrease 

Increase 

Decrease 

Meat  

pe  •  cent 

43 
15 
15 
6 
6 

per  cent 

9 
9 

per  cent 
86 

55 
145 
62 
165 

34 

per  cent 
8 

per  cent 

129 
70 
160 
68 
171 
-17 
25 

Sugar 

Sugar  and  vitamine  J  

Starch 

Starch  and  vitamine|  

Lard..   . 

Lard  and  vitamine  J  

*  The  meat,  sugar,  starch,  and  lard  were  tested  and  found  to  be  free  from 
vitamine  B. 

t  In  this  instance,  the  figure  represents  the  increase  after  60  days,  and  is 
practically  the  same  after  80  days,  since  most  of  the  animals  had  already 
attained  full  size. 

|  Vitamine  given  during  last  55  days. 

The  rats  on  the  protein  diet  did  not  require  the  addition  of  extra  vitamine 
(autolyzed  yeast)  at  all.  This  may  be  regarded  as  the  "sparing  action  of 
protein  on  the  vitamine  requirement."  On  the  other  hand,  the  rats  on  the 
fat  diet  took  the  extra  vitamine  with  great  avidity,  but  showed  only  a  small 
advantage  over  the  controls.  The  replacement  of  some  of  the  fat  by  butter 
was  without  any  significance,  no  improvement  being  noted. 

On  the  starch  diet,  the  rats  actually  needed  extra  vitamine  (about  2  cc. 
per  day)  in  order  to  resume  growth.  This  was  likewise  true  of  the  rats  on 
the  sugar  diet  except  that  they  appeared  not  to  require  as  much  vitamine  for 
growth  as  those  on  the  starch  diet. 


346  THE    VITAMINES 

That  the  protein  content  of  a  food  mixture  is  of  importance,  is 
apparent  from  the  older  work  of  Osborne  and  Mendel,  who  deter- 
mined the  protein  minima  of  various  products  on  rats.  The  experi- 
ments, however,  were  not  carried  out  from  the  viewpoint  of  vit amines. 
In  the  Consideration  of  avian  beriberi  we  have  seen  that  an  excess  of 
carbohydrates  in  the  diet  hastens  the  outbreak  of  the  disease;  an 
excess  of  proteins,  on  the  contrary,  delays  the  onset.  Funk  and 
Dubin  (I.e.  331)  conducted  some  experiments  with  rats,  in  which  it 
is  easier  to  control  the  food  intake.  The  addition  of  vitamine  was 
constant  in  all  diets  and  the  dietary  composition  varied  only  in  its 
content  of  protein,  fat,  starch  and  sugar.  The  composition  of  the 
diets  and  the  results  obtained  are  shown  on  page  345. 

In  another  series  of  experiments,  rats  were  kept  on  a  high  starch 
(curve  VIII,  p.  347)  and  high  meat  (curve  VII,  p.  347)  diet,  without 
any  B-vitamine.  The  rats  on  meat  lived  much  longer  than  those  on 
starch.  The  relationship  may  be  seen  very  well  in  Fig.  64.  Similar 
experiments  were  carried  out  by  Maignon  (1241),  though  he  paid 
no  particular  attention  to  the  vitamines.  Grafe  (1242)  likewise 
made  similar  experiments  on  dogs,  and  Emmett  (1243)  took  up  the 
same  question,  but  his  detailed  report  has  not  yet  appeared.  Experi- 
ments with  food  constituents  mixed  in  unusual  proportions  were  per- 
formed also  by  Osborne  and  Mendel  (1243 a).  Tachau  (1244)  sus- 
pected, several  years  ago,  that  the  utilization  of  certain  dietaiy 
constituents,  such  as  sugar,  may  depend  upon  the  protein  content 
of  the  diet.  As  for  the  significance  of  the  above  data,  they  show 
that  the  protein  concentration  in  a  given  food  mixture  is  of 
greatest  importance.  The  dilution  of  the  protein  and  vitamines  by 
an  excessive  carbohydrate  addition  is  not  without  its  harmful 
sequelae.  In  the  presence  o]  considerable  protein,  the  vitamine  require- 
ments oj  the  organism  are  reduced  to  a  minimum.  We  have  no  tangible 
means  of  explaining  this,  as  yet.  The  protein  may  act  as  "vitamine- 
sparing,"  so  that  less  vitamine  is  required  in  digestion  and  assimila- 
tion, as  compared  with  other  dietary  constituents.  Perhaps  the 
content  of  protein  in  the  new  substance,  mentioned  above,  is  of 
significance  in  this  connection.  From  this,  it  is  clear  that  the  etiology 
of  pellagra  and  war  edema  can  not  be  disposed  of  merely  with  the 
words,  'lack  of  animal  protein."  If  this  were  the  case,  then  we 
should  read  of  many  cases  of  pellagra  in  the  reports  from  Central 
Europe,  which  is,  however,  not  the  case.  Aside  from  this,  it  is 


NUTKITION   IN   MAN 


347 


doubtful  that  two  such  different  diseases  as  pellagra  and  war  edema 
can  arise  from  the  same  cause — lack  of  protein.  At  least,  in  one  case, 
there  must  logically  be  an  error.  Although  we  believe  firmly  in  the 
dietetic  cause  of  both  diseases,  we  cannot  accept,  for  the  present  the 
theory  that  they  arise  from  a  lack  of  animal  protein.  If  this  were  so, 
then  we  ought  to  encounter  much  more  pellagra  among  the  millions 
of  people  that  live  on  an  exclusive  vegetable  diet.  In  both  diseases, 
the  relationship  between  the  proteins,  carbohydrates  and  vitamines 
(perhaps  the  caloric  value  of  the  food  is  often  too  low)  seems  to  be 
incorrect,  and  this  is  all  that  we  know  at  present. 


170 
250 

2.30 


190 


170 

150 
<30 
110 
90 
70 


z 

DAY'S 


FIG.  64.  THE  VITAMINE  REQUIREMENTS   OF   THE   RAT   ON   DIETS   RICH  IN 
PROTEIN,  CARBOHYDRATE  AND  FAT,  RESPECTIVELY 

I,  starch;  V,  starch  +  vitamine  B  after  14  days;  II,  sugar;  VI,  sugar  + 
vitamine  B  after  20  days;  III,  fat;  IV,  meat;  VII,  meat  without  vitamine  B; 
VIII,  starch  without  vitamine  B;  0  =  died;  —>  =  extra  vitamine  B  given 
(Funk-Dubin) 

We  come  now  to  the  consideration  of  the  nutrition  in  Europe  during 
the  war,  and  the  effect  of  this  diet  on  the  general  health  of  the  people. 
The  mode  of  nutrition  in  Germany  at  this  time  has  been  described  by 
Haupt  (1245),  Mason  (1246),  Bornstein  (1247)  and  many  others. 
Zuntz  and  Lowy  (1248)  stated  that  even  at  the  beginning  of  the  war, 
robust  men  lost  about  12  per  cent  of  their  body  weight,  and  Rosenthal 
(1249)  found,  usually,  a  disturbance  of  the  lipoid  metabolism.  The 
influence  on  the  children  and  the  new-born  was  studied  by  Hoffmann 


348  THE   VITAMINES 

(1250),  Jahreiss  (1251)  and  Bloch  (1252)  and  Kaupe  (1252a).  The 
poor  nutrition  evidently  had  no  easily  recognizable  influence  on  the 
intra-uterine  life.  The  size  and  the  weight  of  the  new-born  was  not 
far  different  from  the  normal.  As  regards  the  children  of  school  age, 
there  are  the  reports  of  Abderhalden  (1253),  Pfaundler  (1254)  and 
Blanton  (1255),  all  of  whom  speak  of  a  poor  development  of  the 
children,  both  in  body  and  mind.  In  Belgium,  the  same  observation 
was  made  by  Demoor  (1256)  and  Duthoit  (1256a),  and  in  France 
by  Nobecourt  (1257).  Fronczak  (1258)  reported  on  the  effect  of 
the  war  in  Poland,  and  Rosenfeld  (1259),  in  Denmark  and  Switzer- 
land. 

An  exact  composition  of  the  diet  (per  week)  was  described  by 
Berg  (1260)  in  1917  in  Dresden: 

grams  grains 

Whole  rye  bread 1500      Pork 126 

Wheat  bread 350      Butter  or  margarine 35 

Potatoes 630      Eggs 35 

Kohlrabi 3500      Cheese 14 

Grits 14      Fruits  and  vegetables 400 

Sugar 21      Coffee  substitute .'.  14 

This  gave  a  daily  portion  of  33  grams  protein  (of  which  4  grams 
were  of  animal  origin),  10  grams  fat  and  1217  calories.  We  see  from 
this  that  the  food  was  lacking  in  protein,  fat,  calories,  vitamines  A 
and  (sometimes  apparently)  C.  Many  of  the  observed  dietary  dis- 
turbances in  Germany  were  explained  by  a  poverty  of  fat,  and  Thorns 
(1261)  laid  great  stress  on  this  dietary  component.  It  is  quite  true 
that  a  poverty  of  fat  markedly  reduces  the  caloric  value  of  a  diet, 
but  this  is  frequently  confused  with  a  lack  of  vitamine  A.  For  it  is 
possible  to  live  very  well  without  fat,  as  has  been  shown  again  by 
Osborne  and  Mendel  (1262)  on  rats.  They  demonstrated  that  on  a 
fat-free  mixture,  in  the  presence  of  vitamine  A,  rats  may  grow 
splendidly  and  consume  double  the  quantity  of  food,  as  compared 
with  the  usual  dietary  mixture.  Growth  in  the  first  stage  proceeds 
much  more  favorably,  following  a  greater  protein  intake.  Drum- 
mond  (1262a)  obtained  the  same  results  as  Osborne  and  Mendel  on 
fat-free  food  mixtures.  These  results  find  their  replica  in  experi- 
ments on  children  and  adults,  which  we  have  already  described 

If  the  lack  of  fat  was  not  responsible  for  the  poor  nutrition  in 
Germany,  how  did  nutritive  disturbances  develop?  In  a  series  of 


NUTRITION  IN  MAN  349 

papers,  Hindhede  (1263)  showed  how  this  came  about.  Although  at 
the  beginning  of  the  war,  Germany  was  in  better  position  than  Den- 
mark, as  regards  its  food  reserves,  it  tried  to  conserve  its  cattle  to 
the  very  last,  because  of  the  belief  in  the  special  nutritive  value  of 
animal  protein;  for  this  reason,  the  cattle  consumed  a  large  proportion 
of  the  food  which  could  have  been  utilized  by  the  population.  This 
was  true  particularly  of  pigs.  Kuczynski  and  Zuntz  (1264)  laid  stress 
on  this  point  at  the  time.  In  Denmark,  on  the  contrary,  cattle  con- 
servation was  prevented  by  law,  and  as  a  result  the  Danish  people 
had  a  dietary  composed  as  follows,  according  to  Hindhede  (1265) : 

grams  per  week  grams  per  week 

Whole  rye  bread 1860-2036  Whole  milk 700-2100 

Wheat  bread. 420-  700  Skim  milk 700-1400 

Potatoes 2500^3500  Beef. .  , 175-  500 

Grits 210-  350  Pork 175-  314 

Sugar 448-  467  Butter,  margarine 250 

Cheese 125-  140  Fruits,  vegetables 700 

Beer 700  Coffee 35-60 

This  gave  a  daily  ration  of  57  to  68  grams  protein  (of  which  17  to 
33  grams  were  of  animal  origin),  48  to  59  grams  fat  and  2300  to  2400 
calories.  This  diet  differed  from  the  normal  in  that  it  contained  less 
protein,  and  in  the  substitution  of  vegetable  for  animal  protein. 
What  was  the  effect  of  this  mode  of  nutrition  on  the  health  of  the 
population?  The  mortality  dropped  to  10.4  per  1000,  the  lowest 
figure  of  all  countries.  There  was  a  definite  decrease  of  infections, 
including  tuberculosis,  and  a  notable  drop  in  cancer  mortality,  which 
fell  to  50  per  cent  of  the  previous  figure.  Unfortunately,  it  is  not 
certain  how  much  of  this  improvement  was  due  to  the  general  dietary, 
and  how  much  to  the  abstinence  from  alcohol,  tobacco  and  coffee. 
Of  the  avitaminoses  in  Denmark,  we  have  reported  only  on  oph- 
thalmia, which  we  shall  speak  of  later  on.  In  Germany,  on  the  con- 
trary, although  there  was  a  lack  of  vitamine  A,  ophthalmia  was 
rarely  noted. 

Besides  this,  we  have  observations  on  the  effect  of  the  war  diet 
on  the  general  health  and  the  various  resultant  pathological  occur- 
rences, made  by  Neuhaus  (1266).  In  women,  considerable  war- 
amenorrhea  was  observed  by  Nilsson  (1267).  Puerperal  eclampsia 
was  markedly  decreased  according  to  Gessner  (1268).  Momm 
(1269)  and  Klotz  (1270)  found  that  lactation  was  unfavorably 


350  THE    VITAMINES 

affected.  Zernik  (1271)  noted  a  decrease  in  the  resistance  to  drugs. 
The  number  of  tuberculosis  cases  was  considerably  increased  in 
Germany  (Kieffer,  1272)  and  in  England  (1273).  The  contrary, 
however,  was  true  of  cancer  (Rumpel,  1274)  and  also  of  diabetes, 
according  to  Rumpel,  and  to  Elias  and  Singer  (1275).  The  effect  on 
eye  diseases  was  studied  by  Seefelder  (1276). 

It  is  evident  from  what  has  been  said,  that  in  spite  of  the  small 
amount  of  animal  protein  in  the  war  diet  of  Central  Europe,  no  out- 
break of  pellagra  occurred;  this  was  also  commented  on  by  S.  Harris 
(1277).  In  addition,  despite  the  lack  of  vitamine  A,  very  little 
ophthalmia  prevailed.  On  the  other  hand,  there  was  considerable 
scurvy  and  hunger  edema.  We  see  from  the  well-controlled  diet  of 
the  Danish  population  during  the  war,  that  68  grams  protein  (con- 
taining 27  gm.  animal  protein)  per  day,  50  grams  fat  and  2500 
calories  completely  sufficed  to  cover  the  requirements.  Hindhede's 
experiments  show  that  the  amount  of  protein  might  even  be  decreased 
further,  and  could  be  substituted  by  good  vegetable  protein. 


CHAPTER  VI 

PELLAGRA 

Pellagra  is  a  non-contagious,  endemic  disease  of  corn-eating  popu- 
lations, prevalent  in  northern  Italy,  Rumania,  southern  Tyrol  and 
North  America.  The  disease  consists  in  a  specific  erythema  of  the 
skin,  stomatitis,  gastro-enteritis  and  profound  degenerative  changes 
of  the  central  nervous  system.  The  progress  is  acute  or  chronic, 
and  the  mortality  is  appreciable.  Pellagra  has  been  described  by 
Bouchard  (1278),  Roussel  (1279),  Marie  (1280),  Roberts  (1281), 
Schilling  (1282),  Niles  (1283)  and  Harris  (1284). 

GEOGRAPHICAL   DISTRIBUTION   OF   PELLAGRA 

The  greatest  prevalence  of  this  disease  is  in  northern  Italy,  where 
it  has  been  known  since  1700,  affecting  Piemont,  Lombardy,  Venice 
and  Emilia.  Central  Italy  is  much  less  affected  while  the  south  is 
almost  free  of  the  disease.  It  is  unknown  in  Corsica,  Sardinia  and 
Sicily.  Pellagra  in  Italy  is  evidently  decreasing;  in  1898,  there 
were  3987  fatal  cases,  while  in  1905,  only  2359.  The  census  shown 
as  follows : 

Year  Cases  of  Pellagra 

1879 97,855 

1881 104,067 

1899 72,603 

1905 55,029 

1910 33,869 

In  Austria,  especially  in-  southern  Tyrol,  numerous  cases  were 
recognized  between  1875  and  1905;  since  this  time,  however,  they 
have  diminshed  so  rapidly  that  the  number  of  cases  in  Rovereto, 
according  to  Weiss  (1285),  dropped  from  8053  in  1904  to  3503  in  1912, 
upon  the  introduction  of  potatoes.  In  Roumania,  the  number  of 
pellagra  patients  were  calculated  at  about  75,000  a  few  years  ago; 
the  disease  seems  to  have  increased  in  Serbia  and  Bulgaria.  In 
England,  some  cases  have  been  described  by  Sambon  and  Chalmers 
(1286),  Box  (1287)  and  Low  and  Yellowlees  (1288).  Still,  the 
diagnosis  of  sporadic  cases  seems  to  be  somewhat  uncertain,  though 

351 


352  THE    V  IT  AMINES 

not  impossible.  In  France,  the  disease  is  supposed  to  have  been 
recognized  only»since  the  time  of  Napoleon  I,  especially  in  the 
swampy  province  of  Landes,  but  it  has  been  entirely  eradicated  by 
Roussel  (I.e.  1279). 

In  Egypt,  especially  in  Lower  Egypt,  we  find  an  insignificant 
number  of  pellagra  cases.  In  Canada,  cases  were  observed  by 
Pinault  (1289)  and  Rolph  (1290).  In  North  America,  till  a  few 
years  ago,  pellagra  had  been  steadily  increasing,  apparently  since 
1880;  some  investigators  believe,  on  the  contrary,  that  it  appeared 
only  in  1900.  The  progress  of  the  disease  here  is  much  more  acute 
than  in  Italy,  and  the  mortality  markedly  greater.  The  endemia 
is  most  severe  in  the  southern  states  (especially  Texas,  Arkansas, 
Louisiana,  Mississippi,  Kentucky,  Tennessee,  Alabama,  Virginia, 
North  and  South  Carolina,  Georgia)  and  in  Mexico.  Lavinder 
(1291),  between  1907  and  1921,  counted  30,000  pellagra  cases, 
with  40  per  cent  mortality.  Petersen  (1292)  reported  recently  that 
in  the  United  States,  during  1915,  there  were  10,663  fatal  cases; 
in  1916,  there  were  6289.  During  this  time,  in  the  southern  states, 
the  pellagra  cases  were  supposed  to  represent  about  0.5  per  cent  of 
the  population,  which  would  make  about  165,000  cases.  In  the 
Canal  Zone,  cases  were  reported  by  Deeks  (1293),  arising  on  a 
carbohydrate-rich  diet.  Tuttle  (1294)  has  described  cases  in  South 
America.  McDonald  (1295)  observed  pellagra  in  Antigua;  Nicholls 
(1296),  on  the  Island  of  St.  Lucia;  de  Kock  and  Bonne  (1297),  in 
Surinam. 

PROGRESS   OF   PELLAGRA 

Pellagra  spreads  in  rural  districts,  while  the  cities  remain  free 
from  the  disease.  In  Italy  and  Egypt,  the  disease  attacked  the 
poorer  population;  in  North  America,  however,  even  well-to-do 
farmers  who  follow  the  usual  mode  of  country  life  are  not  immune. 
According  to  Roberts  (1298),  the  picture  of  pellagra  has  changed 
in  the  United  States  in  recent  years;  here,  the  acute  form  resembles 
the  Italian  type,  and  some  forms  are  so  mild  tthat  they  may  be 
easily  overlooked.  The  disease  occurs  more  in  women  than  in  men; 
in  fact,  the  ratio  between  both  sexes  is  given  variously  as  2  : 1  or  3  : 1. 
According  to  Siler  and  Garrison  (1299)  and  Grimm  (1300),  pellagra 
often  appears  after  a  birth  and  in  83  per  cent  of  all  cases,  it  breaks 
out  in  families  in  straitened  circumstances. 


PELLAGRA  353 

The  first  attacks  and  relapses  appear  in  Italy  mostly  early  in  the 
year — March  or  April;  in  North  America,  beginning  in  February 
and  lasting  throughout  the  summer  into  late  autumn.  Wellman  and 
Sparkes  (1301)  have  reported  some  winter  cases. 

Pellagra  is  no  respecter  of  age,  but  it  is  seldom  seen  in  infants. 
Niles  (I.e.  1283),  for  example,  says  that  he  has  personally  never  seen 
a  case  under  5  years  and  very  few  under  10  years  of  age.  According 
to  Snyder  (1302),  the  disease  is  often  seen  in  older  children;  thus, 
10  per  cent  of  all  cases  appear  before  the  age  of  15.  In  one  case  of 
a  diseased  mother  who  nursed  her  child  for  6  weeks,  pellagra 
symptoms  appeared  in  the  child  a  few  days  later  on  an  artificial 
diet.  Weston  (1303)  found  in  Columbia  15  cases  among  children, 
one  of  which  had  a  sick  mother;  the  child  developed  pellagra 
shortly  after  being  weaned.  Voegtlin  and  Harries  (1304)  reported 
the  very  interesting  case  of  a  5  months  old  breast-fed  baby,  whose 
mother  showed  no  symptoms.  Byfield  (1305)  described  a  pellagra- 
like  disease  in  17  children  under  4  years  of  age,  for  which,  however, 
he  assumed  an  infectious  and  not  an  alimentary  cause.  According 
to  Murphy  (1306),  the  disease  is  much  milder  in  children  and  is 
without  nervous  symptoms. 

Pellagra  is  not  hereditary,  but  frequently  the  children  of  pellagrans 
show  stunted  growth,  mental  weakness  and  other  signs  of  degener- 
ation. It  is  not  contagious ;  never  was  there  an  infection  in  an  orphan 
asylum  or  among  the  guests  of  a  health  resort;  according  to  Gold- 
berger  (1307),  there  was  never  any  pellagra  among  physicans  and 
nurses  of  a  pellagra  hospital. 

The  course  of  the  disease  varies;  there  are  severe  acute  cases 
terminating  fatally  in  a  few  weeks,  and  chronic  types,  with  recoveries 
and  relapses,  of  many  years  duration.  In  an  examination  of  100 
cases,  Wood  (1308)  noted  the  first  symptoms,  divided  as  follows: 
skin,  100  per  cent;  gastro-intestinal  disturbance,  77  per  cent; 
mouth  symptoms  appearing  sometimes  before  the  skin  symptoms. 
Of  the  latter,  97  per  cent  appeared  on  the  hands  and  the  forearms, 
and  in  39  cases,  on  the  hands  only.  The  uncovered  parts  of  the  body 
are  involved,  for  the  most  part,  but  in  one  case,  the  anus,  and  in 
two  cases,  the  back  was  affected.  According  to  Roberts  (I.e.  1281), 
there  are  altogether  four  different  types  of  the  disease. 


354  THE   VITAMINES 

1.  Acute,  malignant  form 

Duration  is  from  two  weeks  to  three  months.  It  may  appear  as  the 
first  attack  or  during  the  progress  of  the  disease,  or  end  in  chronic 
cases  with  prostration,  convulsions,  fever  and  diarrhea.  Contin- 
uous fever  (38  to  40.5°C.);  pulse  120  to  130,  small  and  arrythmic; 
tongue,  deep  red,  fissured  and  painful;  painful  stomatitis  and 
pharyngitis;  nausea;  typical  dermatitis;  skin  petechiae.  In  addi- 
tion there  are  frequently  tremors,  convulsions,  tetanus,  delirium  and 
incontinence.  This  type,  is  often  called  "typhoid  pellagra." 

2.  Light  sub-chronic  jorm 

Prevalent  in  young  individuals;  light  cases  with  dermatosis  and 
dyspepsia,  disappearing  when  cured.  Duration,  1  to  2  years;  no 
fever.  This  form  is  very  common  in  Italy  and  Roumania;  less  so 
in  America.  The  symptoms  consist  of  a  light  dermatosis  on  the  backs 
of  the  hands  and  wrists,  reddening  of  the  mouth  mucosa,  consti- 
pation or  slight  diarrhea. 

3.  Severe,  cachectic  sub-chronic  form 

Temperature  at  the  beginning,  37.7°C;  pulse,  100.  Very  severe 
symptoms  along  the  digestive  tract;  the  tongue,  deep  red,  eroded; 
mouth  and  pharynx  mucosa,  deep  red  and  painful;  gums,  swollen 
and  bleeding;  nausea,  vomiting,  gastralgia,  persistent  serous  diarrhea, 
loss  of  weight.  Later,  there  are  cerebral  symptoms,  mental  weak- 
ness and  skeleton-like  emaciation. 

4.  Chronic  form 

Duration,  1  to  20  years.  In  most  cases,  three  periods  are  differ- 
entiated. A)  First  period,  with  dermatosis  and  dyspepsia,  as  in 
light  sub-chronic  cases.  B)  Second  period,  symptoms  along  the 
digestive  tract. become  more  severe;  gastralgia,  tabes-like  stomach 
crises,  vomiting,  diarrhea  and  tenesmus  appears.  Pulse,  80  to  100, 
with  dyspepsia  and  light  dropsy.  Typical  dermatosis;  on  recur- 
rence the  skin  of  the  feet  is  brownish  red  and  scaly ;  backs  of  the  hands 
are  brownish  red,  wrinkled  and  aged.  To  this  picture  there  are 
added  brain  and  spinal  cord  symptoms;  uncertain  walk,  vertigo, 
tremor,  epileptic  and  tetanic  attacks,  contractures,  accentuated 


PELLAGRA  355 

reflexes.  C)  Sooner  or  later  in  the  third  period,  there  is  cachexia 
and  mental  weakness,  with  hemiplegia  or  paraplegia,  and  finally 
with  serous  diarrhea,  petechiae,  muscle  atrophy  and  incontinence. 
Normal  temperature  prevails,  as  a  rule ;  only  exceptionally  is  terminal 
fever  observed  in  the  last  days  of  the  disease.  In  addition  to  these 
four  types,  there  are  light  abortive  forms,  which  we  shall  describe 
in  the  consideration  of  expermental  pellagra  in  man. 

SYMPTOMATOLOGY  AND  PATHOLOGY  OF  PELLAGRA 

1.  G astro-intestinal  tract 

The  pellagrous  tongue  is  of  diagnostic  importance:  in  the  early 
stage,  coated;  later,  deep  red,  fissured  in  the  middle  and  around 
the  edge,  often  painful.  The  gums,  spongy,  red,  easily  bleeding  as 
in  scurvy;  sometimes  there  is  alveolar  pyorrhea.  Swelling  and  red- 
dening of  the  mouth  and  pharynx  muscosa  with  aphthae  or  small 
vesiculae,  pyrosis,  gastralgia,  nausea  and  vomiting  occur.  In  an 
investigation  of  the  stomach  of  20  cases,  Johnson  (1309)  found  a 
lack  of  hydrochloric  acid  in  16.  This  finding  was  confirmed  by 
Givens  (1310),  who  noted,  however,  cases  in  which  hydrochloric 
acid  and  pepsin  were  present,  especially  in  children.  Pellagrous 
diarrhea  is  of  importance;  in  a  spring  attack  it  may  happen  that 
there  will  be  from  10  to  20  evacuations  daily,  often  slimy  and  bloody. 
In  the  final  stages,  the  evacuations  occur  still  more  frequently,  serous 
and  colorless;  indicanuria  is  often  noted.  All  the  symptoms  along 
the  gastro-intestinal  tract  appear  to  be  of  central  nervous  origin. 
Siler  (1311)  preferred  not  to  regard  as  pellagra  those  cases  without 
gastro-intestinal  disturbances,  though  it  seems  to  us  that  such  a 
differentiation  would  be  only  artificial. 

In  acute  cases,  in  the  stomach,  swelling  and  reddening  with  erosions, 
especially  at  the  pylorus  were  found.  In  chronic  cases,  the  mucous 
membrane  of  the  stomach  was  pale,  covered  with  mucous,  and  the 
muscles  were  atrophic.  In  the  intestine,  in  acute  cases,  there  was 
enteritis  with  ulcerations  in  the  small  and  large  intestine,  less 
frequently  in  the  duodenum;  in  chronic  cases,  according  to  Lynch 
(1312),  the  mucosa  was  pale,  covered  with  mucus,  and  atrophic; 
the  musculosa  was  thinned.  The  liver  was  tough,  atrophic,  and 
occasionally  showed  fatty  degeneration. 


356 


THE   VITAMINES 


FIG.  65.  TYPICAL  PELLAGRA  ERYTHEMA 
ON  THE  BACK  OF  THE  NECK  (ROBERTS) 


FIG.  66.  PELLAGRA  IN  A  NEGRESS 
(ROBERTS) 


FIG.  67.  PELLAGRA  (ZELLER) 


PELLAGRA  357 

2.  Skin 

The  pellagrous  dermatitis  is  one  of  the  most  important  of  the  early 
symptoms.  In  light  forms,  this  dermatitis  is  maculo-papular;  in 
severe  acute  types,  vesicular  with  intensive  reddening,  edema  and 
pain.  The  dermatitis  appears  in  a  typical  manner  symmetrically 
on  the  backs  of  the  hands,  and  wrists  (the  pellagrous  glove),  then 
on  the  neck,  less  often  on  the  face,  and  on  the  feet  (the  pellagrous 
shoe).  This  dermatitis  was  likened  by  Gurd  (1313)  to  X-ray 
dermatitis.  The  pellagrous  glove  and  shoe  extends  frequently 
one-third  of  the  way  up  the  forearm  and  leg  respectively.  On  the 
face,  the  dermatitis  appears  on  the  nostrils,  temples  and  behind  the 
ears;  sometimes,  ecchymoses  appear  on  the  eye-lids.  On  the  elbows 
and  the  forearm  the  skin  remains  rough  and  scaly  for  a  long  time; 
this  protracted  scaling  may  appear  on  the  face,  shoulders  and,  in 
fact,  on  the  whole  body.  In  chronic  cases,  the  nails  sometimes 
become  grayish  white,  thickened  and  brittle.  On  recovery,  the  skin 
appears  brownish  red  and  dark  brown  on  the  affected  places;  in 
rare  cases,  this  extends  to  the  entire  body. 

It  is  of  etiologic  importance  that  in  the  production  of  human 
experimental  pellagra  by  Goldberger  (1314),  the  first  symptoms 
appeared  on  the  scrotum.  This  condition  has  inclined  some  investi- 
gators to  doubt  the  validity  of  Goldberger's  findings,  pointing  out 
that  it  had  nothing  to  do  with  real  pellagra.  Because  of  this,  it  is 
important  to  state  here  that  Deiaco  (1315),  Merk  (1316),  Deeks 
(I.e.  1293),  Crosby  (1317)  and  Wood  (I.e.  1308)  observed  this 
localization  of  the  dermatitis  as  the  first  sign  in  their  cases. 

This  specific  dermatitis  appears  to  be  of  trophoneurotic  central 
origin,  but  the  influence  of  the  sun  rays  cannot  be  altogether  denied. 
The  burning  of  the  hands  and  feet,  a  very  disagreeable  occurrence 
independent  of  the  dermatitis,  belongs  to  the  nervous  symptoms. 

3.  The  nervous  system 

Pain  in  the  back  is  one  of  the  early  symptoms  of  severe  cases.  The 
reflexes  are  accentuated.  The  muscles  become  atrophic  in  half  of 
the  chronic  cases,  with  fatty  degeneration;  contractions  in  a  flexed 
position  sometimes  develop  in  the  hands  and  feet.  The  degeneration 
reaction  is  usually  lacking.  Among  the  later  symptoms,  are  tremor, 
especially  of  the  hands,  infrequently  of  the  tongue  and  lips,  and 


358 


THE   VITAMINES 


cramps  of  some  muscle  groups;  in  severe  cases,  there  is  spastic  or 
atactic  gait.  As  terminal  symptoms,  there  are  tetanic,  epileptic 
attacks,  and  paralysis  of  the  sphincters.  Mental  disorders  belong 
to  the  typical  picture  of  severe  pellagra.  The  initial  symptoms  are 
insomnia,  quietness,  sadness,  later  hypochondria  and  finally  psychoses 


FIG.  68.  SPINAL  CORD;  LUMBAR 
REGION  (Moir) 


FIG.  69.  CELLS  OF  VENTRAL 
HORN  WITH  ASYMMETRIC 
NUCLEI  (MOTT) 


FIG.  70.  SCIATIC  NERVE  IN 
PELLAGRA;  LONGITUDINAL 
SECTION   (MOTT) 


• 


FIG.  71.  CELL  OF  BETZ  IN 
MOTOR  CORTEX  (MOTT) 


of  all  kinds.  The  latter  develop  in  10  per  cent  of  all  cases  in  Italy, 
in  about  5  per  cent  in  America,  apparently  more  often  in  the  chronic 
form.  According  to  Sandy  (1319)  the  number  of  pellagra  psychoses 
among  the  inmates  of  insane  asylums  is  very  large  in  the  southern 
states.  Miller  and  Ismail  (1320)  investigated  the  various  types  of 
pellagrous  lunatics  in  Egypt,  classifying  750  cases  as  follows: 


PELLAGKA  359 

Amentia  (42  per  cent — typical  first  stage) 320 

Mania 115 

Melancholia 113 

Dementia 103 

The  mortality  was  32  per  cent;  aside  from  this,  47  per  cent  were 
discharged,  with  unknown  issue. 

In  the  brain,  the  following  lesions  were  found :  pia  and  arachnoid 
were  thickened,  with  a  milky  cloudiness  and  ecchymoses.  In  the 
brain,  mostly  edema  and  hyperemia  with  hydrops  of  the  ventricles 
are  noted.  In  chronic  cases,  the  brain  and  the  brain  convolutions, 
especially  the  frontal,  frequently  become  atrophic,  hard  and  anemic. 
Miscrocopically,  cortical  nerve  cells  are  degenerated,  with  swelling, 
vacuole  formation,  displacement  of  the  swollen  nucleus  to  the  side, 
and  later  atrophy  of  the  degenerated  cells. 

In  the  spinal  cord,  in  acute  cases,  dilatation  of  the  blood  vessels 
and  edema  are  found;  in  chronic  cases,  chiefly  degeneration  of  the 
dorsal  fibers  and  the  direct  pyramidal  tract.  Scattered  localities 
with  disappearance  of  the  nerve  fibers  in  the  whole  white  substance 
of  the  spinal  cord  were  also  noted.  With  this,  the  dorsal  roots  are 
also  degenerated  with  arterial  thickening.  In  the  gray  substance, 
and  also  in  the  cells  of  the  ventral  and  dorsal  horns,  we  find  pigmen- 
tation, swelling  of  the  cell  protoplasm,  chromatolysis  and  displace- 
ment of  the  nucles  to  one  side.  In  the  sympathetic  nerve,  especially 
in  the  abdominal  ganglia,  degeneration  was  observed.  It  must  be 
explicitly  stated  that  in  the  nerve  centers,  there  was  not  a  trace  of 
inflammatory  lesions;  all  the  changes,  according  to  Mott  (I.e.  890), 
are  exclusively  of  a  degenerative  nature. 

Microscopic  hemorrhages  were  often  noted  in  the  nerve  centers. 
In  one  of  Chalmer's  (1321)  cases  in  Roumania,  the  cause  of  death 
was  a  marked  hemorrhage  in  the  lower  cervical  and  dorsal  part  of 
the  spinal  cord.  In  the  sciatic  nerve,  Mott  found  scattered  degener- 
ation of  nerve  fibers. 

4.  Circulatory  system 

In  the  blood,  according  to  Findlay  (1318),  an  increase  in  lympho- 
cytes is  mostly  found.  Hemoglobin,  about  80  per  cent;  pulse,  usually 
100  and  in  acute  pellagra,  up  to  120;  blood-pressure  is  mostly  low. 
Examination  of  the  blood  for  microorganisms  was  negative.  Bardin 
(1322)  found  an  increase  in  the  number  of  small  and  large  lympho- 


360  THE   VITAMINES 

cytes  and  a  decrease  in  polymorphonuclear  neutrophiles.  Micro- 
scopic findings  were  recorded  by  Nagamatsu  (1323).  Examination 
of  the  cerebrospinal  fluid,  according  to  Lorenz  (1324),  gave  no  reason 
for  regarding  pellagra  as  an  infection  of  the  central  nervous  system. 
The  heart  is  mostly  atrophic,  pigmented,  fragile  and  occasionally 
shows  fatty  degeneration.  Sometimes  there  is  lung  edema,  hyper- 
emia  and  hydro  thorax. 


i 


V         v 


FIG.  72.  RARIFICATION  OF  THE  PHALYNGEAL  ENDS  IN  PELLAGRA  (ROBERTS) 

5.  Bones 

Lombroso  (1325)  and  Babes  and  Sion  (1326)  observed,  in  some 
cases,  brittleness  of  the  ribs  and  long  bones.  Fractures  of  the  long 
bones  were  also  noted.  Similar  lesions  are  characteristic  of  human 
and  experimental  scurvy.  Roberts  (I.e.  1281)  gives  some  X-ray 
pictures  of  pellagrous  bones,  showing  a  rarification  of  the  bone  ends. 

6.  Sexual  organs 

In  acute  cases,  vulvo-vaginitis  is  not  infrequently  observed,  and 
in  rare  cases,  there  is  gangrene  of  the  labia.  In  chronic  cases,  amen- 
orrhea  frequently  occurs.  Abortion  was  noted  in  20  per  cent  of 


PELLAGBA  361 

pcllagrous  pregnant  women;  hemorrhages  following  birth  are  not 
uncommon.  In  latent  cases,  following  pregnancy,  an  acute  condition 
is  not  infrequently  brought  on;  similar  observations  have  been 
made  in  beriberi. 

7.  Other  organs 

Regarding  eye  symptoms,  Marie  (I.e.  1280)  described  conjuncti- 
vitis with  pterygium  and  hemeralopia,  and  sometimes  pigmentary 
retinitis.  Calhoun  (1327)  found  that  certain  visual  disturbances, 
especially  certain  forms  of  color  blindness,  precede  other  symptoms 
of  pellagra,  so  that  they  may  be  used  in  the  diagnosis. 

As  regards  the  endocrine  glands,  Beeson  (1328)  observed,  out  of 
316  cases,  25  cases  of  thyroid  complications.  Modinos  (1329)  noted 
cases  in  Egypt  with  enlarged  adrenals,  about  twice  the  usual  size. 
In  cases  observed  by  Wilson  (1330),  also  in  Egypt,  these  glands  were 
much  lighter  in  weight.  Morse  (1331)  found  severe  atrophic  degen- 
erative changes  in  the  thyroid  and  the  adrenal  medulla.  In  the 
latest  reports,  especially  from  Egypt,  special  attention  was  paid  to 
the  insufficiency  of  the  adrenals  (Wilson,  I.e.  1330). 

CHEMICAL   PATHOLOGY 

Koch  and  Voegtlin  (1332)  analyzed  the  spinal  cord  and  brain  of 
pellagrins  and  found  that  the  figures  agreed,  in  general,  with  those 
obtained  in  pigeon  beriberi.  One  of  the  most  important  findings 
was  the  loss  of  lipoids,  particularly  in  the  spinal  cord. 

Indicanuria  was  found  by  Ridlon  (1333)  in  90  .per  cent  of  his 
cases.  Murlin  (1334)  found  high  amino  nitrogen  and  hippuric  acid 
values  (two  to  three  times  greater  than  normal),  indicating  a  poorer 
nitrogen  utilization.  If  the  carbohydrate  diet  is  replaced  by  a  protein 
diet,  then  the  values  found  are  smaller.  The  effect  of  both  of  these 
diets  on  the  composition  of  the  blood  was  investigated  by  Lewis 
(1335).  The  carbohydrate  diet  gave  smaller  urea  and  non-protein 
nitrogen  figures,  but  otherwise  the  findings  were  normal.  Jobling 
and  Maxwell  (1336),  as  well  as  Sullivan  and  Stanton  (1337),  deter- 
mined the  alkali  reserve  of  the  blood ;  the  figures  varied  only  slightly 
from  the  normal;  no  acidosis  was  noted.  Sullivan  and  Jones  (1338) 
found  indican  in  the  saliva  in  one  case;  in  addition,  there  was  a 
thiocyanide  reaction,  indicating  a  retardation  of  protein  metabolism. 


362  THE   VITAMINES 

METABOLISM 

The  first  exact  metabolism  experiments  in  pellagra  were  made  by 
Myers  and  Fine  (1339).  The  milk-vegetable  diet  used  was  well 
utilized.  Nicolaidi  (1340)  observed  large  food  losses  in  the  stool. 
Albertoni  and  Tullio  (1341)  obtained  a  negative  nitrogen  balance  on 
corn;  the  balance  became  positive  on  meat.  Hunter,  Givens  and 
Lewis  (1342)  found  in  seven  cases  of  pellagra  a  positive  nitrogen 
balance  on  a  pellagra-producing  diet.  Boyd  (1343)  found,  in  a 
metabolism  experiment  on  Turkish  prisoners  in  Egypt,  in  confirma- 
tion of  earlier  findings,  a  decrease  of  hydrochloric  acid  in  the  stomach, 
diminished  pancreatic  secretion,  protein  and  fat  loss  in  the  stool 
(remarkable  in  a  disease  caused  by  a  lack  of  protein)  and  a  very  poor 
protein  assimilation.  The  latter  is  especially  emphasized  in  recent 
publications  (Sullivan,  Stanton  and  Dawson,  1343a) . 

PROGNOSIS 

In  Italy,  55,029  cases  of  pellagra  were  recorded  in  1905,  with  2359 
deaths,  equivalent  to  more  than  4  per  cent  mortality.  In  America, 
on  the  contrary,  the  prognosis  is  much  more  serious,  namely,  50  per 
cent  death  rate  in  the  asylums  and  20  to  25  per  cent  in  private 
practice.  It  is  obvious  that  the  picture  of  pellagra,  because  of  the 
dietetic  factor,  frequently  varies  with  the  well-being  of  the  popula- 
tion, assuming  either  a  mild  or  more  severe  type. 

THE   RELATIONSHIP  OF  PELLAGRA  TO  THE  ACCEPTED  AVITAMINOSES — 
BERIBERI   AND    SCURVY 

In  the  short  paragraph  on  the  relation  of  beriberi  to  scurvy  in 
South  Africa  (p.  296),  we  have  seen  that  scurvy  develops  on  a  diet 
consisting  of  very  little  animal  protein  and  corn,  without  a  sign  of 
pellagra.  This  is  true  also  of  a  widespread  occurrence  of  scurvy, 
which  G.  R.  Hopkins  (I.e.  939)  described  in  a  population  which  fed 
itself  almost  exclusively  on  corn.  Marie  (I.e.  1280)  and  Viswallingam 
(1344)  looked  upon  scurvy  as  a  predisposing  cause  of  pellagra. 

As  for  the  relation  of  pellagra  to  beriberi,  Sheppard  (1345)  saw 
pellagra  in  Singapore  on  an  exclusive  rice  diet.  Stannus  (1346) 
stated  that  he  saw  a  number  of  cases  in  Nyasaland  on  rice  (only 
partially  husked) ;  vegetables,  fish  or  meat  were  eaten  only  once  in 
14  days.  The  disease  progressed  without  any  characteristic  exan- 


PELLAGRA  363 

thema,  although  certain  signs  of  the  skin  affection  were  visible 
around  the  mouth.  On  the  other  hand,  a  pellagra-like  erythema 
was  observed  in  beriberi  by  Schiiffner  and  Kuenen  (1347).  Night- 
ingale (1348)  saw  1210  pellagra-like  cases,  in  South  Rhodesia,  which 
he  called  "zeism."  The  disease  developed  on  milled  corn,  was 
mild  and  without  exanthema,  but  with  light  dermatitis  and  mouth 
symtoms.  He  stated  that  cod  liver  oil  therapy  was  quite  favorable. 
Edwards  (1349)  designated  as  "peripheral  neuritis"  a  disease  met 
with  in  Jamaica  among  the  poor  inhabitants  who  live  on  a  nitrogen- 
poor  diet.  The  symptoms  resemble  pellagra  more  than  beriberi. 
Finally,  we  wish  to  comment  upon  the  composition  of  the  diet,  which 
Braddon  (I.e.  866)  regarded  as  productive  of  beriberi.  It  was 
exceptionally  poor  in  animal  protein  and  must  have  lead  to  pellagra, 
according  to  the  newest  conceptions.  This  short  chapter  was 
introduced  here,  not  so  much  to  demonstrate  the  relationship  between 
pellagra  and  the  avitaminoses,  but  to  show  that  we  may  be  dealing 
with  mixed  forms  under  certain  conditions,  which  indicates  perhaps 
that  if  the  diet  is  lacking  in  one  constituent,  it  is  likely  that  others 
may  be  lacking  at  the  same  time. 

MODE    OF   DEVELOPMENT   OF   PELLAGRA 

Although  theoretically  pellagra  may  also  develop  without  any 
corn  consumption,  it  appears  and  disappears,  in  practice,  with  corn 
cultivation.  Thus,  corn  was  introduced  into  Egypt  in  1840,  and  in 
1847,  the  first  cases  of  pellagra  were  noted.  In  Spain,  the  disease 
has  been  endemic  for  two  centuries,  having  appeared,  there  with  the 
beginning  of  corn  cultivation;  at  present,  considerable  rye,  wheat  and 
oats,  but  very  little  corn,  are  grown  and  the  disease  has  almost  been 
stamped  out  there.  Weiss  (1350)  noted  that  since  1905,  after  the 
exclusion  of  corn  from  the  diet,  cases  in  the  Tyrol  appeared  more 
rarely.  Alpago-Novello  (1351),  in  an  Italian  province,  partially  sub- 
stituted corn  growing  by  turnips  and  potatoes,  whereupon  the  number 
of  cases  decreased.  It  is  likewise  not  without  significance  whether 
hand  ground  or  machine  milled  corn  is  used.  In  the  United  States, 
it  has  often  been  found  that  in  the  mountainous  districts  where  only 
the  hand  milled  variety  is  used  no  cases  of  pellagra  occur;  the  con- 
trary is  true  in  small  industrial  cities  where  the  natives  use  prepared 
cornmeal,  as  brought  out  by  Wood  (1352).  Blosser  (1353)  observed 


364 


THE    VITAMINES 


130  cases  develop  on  a  diet  containing  a  considerable  amount  of 
cane  sugar  products,  especially  molasses.  By  omitting  the  latter, 
which  is  greatly  relished  in  the  southern  states,  an  improvement  was 
noted.  That  a  diet  very  rich  in  carbohydrates  is  consumed  in  dis- 
tricts where  pellagra  prevails,  is  evident  from  the  report  of  Jobling 
and  Petersen  (1354).  Babes  (1355)  remarked,  advisedly,  that  when 
in  February  1918,  the  entire  population  of  Bucharest  was  placed  upon 
bread  made  of  corn  and  wheat,  numerous  cases  of  pellagra  appeared 
as  early  as  in  May,  so  that  three  months  were  necessary  for  the 
development  of  the  disease.  Lombroso  (I.e.  1325)  and  Camurri  (1356) 
describe  a  dietary  composition  in  the  Italian  pellagra  districts : 

Lombroso  Camurri 

Grams  daily  Grams  daily 

Corn 1091   Polenta 1500 

Rice  and  barley 67   Rice 100 

Beans 60   Beans 100 

Potatoes 67   Potatoes 100 

Vegetables 250   Vegetables 100 

Lard 21   Lard 20 

Olive  oil 33      Olive  oil 10 

Fish 67      Milk 100 

Poultry. 27      Cheese 50 

Apart  from  this,  great  differences  were  noted  in  the  summer  and 
winter  diets,  which  explains  very  well  the  periodic  occurrence  of 
pellagra  in  spring.  It  seems  that  the  Italian  population  feeds  itself 
very  poorly  in  the  winter.  Lombroso  reported  on  such  dietary 
changes  in  the  province  of  Ferrara: 


DIET  IN  8 
WINTER  MONTHS 

DIET  IN  4 
SUMMER  MONTHS 

Polenta                                          .    .           ... 

grams  per  day 
1000 

grams  per  day 

160 

Milk  ..     . 

Eggs  

Almost  none 

Almost  none 

Onions  

One  daily 

Two  daily 

Corn  bread 

50 

400 

Home-made  bread  

50 

200 

Meat.  .                                                 

10 

60 

Cheese  

5 

20 

Beans  

150 

40 

Fish  

20 

Very  little 

PELLAGRA  365 

These  figures  were  corroborated  by  Devoto  (1357).  Wilson  (1358) 
saw  the  development  of  pellagra  on  a  diet  containing  92  grams 
protein  and  2200  calories.  The  protein  consisted  of  J  wheat  and 
|  corn;  10  per  cent  of  the  war  prisoners  developed  pellagra  in  one 
year,  the  disease  appearing  only  after  hard  labor.  Boyd  and  Lelean 
(1359)  observed  pellagra  among  6000  Turkish  prisoners,  to  the  extent 
of  18  per  cent,  while  among  German  prisoners,  there  were  no  cases; 
it  was  only  when  they  were  assigned  hard  labor  to  perform  that  65 
cases  appeared,  according  to  Lelean  (1360).  Bouchard  (I.e.  1278) 
noted  that  shepherds  developed  pellagra,  while  cow  attendants 
remained  free  from  the  disease.  The  difference  was  supposed  to  lie 
in  the  milk  consumed. 

The  mode  of  occurrence  of  pellagra  was  studied  very  accurately  by 
Goldberger  and  his  co-workers.  Goldberger,  Wheeler  and  Syden- 
stricker  (1361)  compared  the  diets  of  diseased  and  healthy  natives  in 
a  small  industrial  city  in  South  Carolina.  The  chief  difference  was 
in  the  milk,  meat,  vegetable  and  fruit  consumption.  The  amount 
of  protein  was  about  85  grams,  of  which  one-third  was  of  animal 
origin.  The  reason  why  other  investigators  did  not  observe  the  above 
differences  is  because  the  pre-pellagrous  diet,  and  not  that  which  is 
consumed  at  the  time  of  the  attack,  must  be  studied ;  for  in  this  case, 
the  disease  is  already  improving.  Sydenstricker  (1362)  could  show 
by  statistical  data,  that  the  cases  become  more  numerous,  when  the 
people,  because  of  increased  prices,  must  limit  their  expenditures. 
Apparently,  the  decrease  in  the  number  of  pellagra  cases  in  the  United 
States  during  the  war  is  due  to  the  great  prosperity  of  the  population. 
Goldberger  and  Wheeler  (I.e.  109)  produced  experimental  pellagra  in 
11  prisoners.  The  diet,  accurately  controlled,  was  analyzed  by 
Sullivan  and  Jones  (1363),  and  the  vitamine  content  was  tested  on 
animals  by  Sullivan  (1364).  The  diet  consisted  of  41  to  54  grams 
protein,  of  which  80  to  97  per  cent  was  of  vegetable  origin,  and  it 
seemed  to  be  poor  in  vitamines  B  and  A.  The  controls  received  a 
better  diet.  The  experiment  lasted  6|  months,  during  which  time, 
out  of  11-  experimental  subjects,  5  developed  symptoms,  which 
must  be  regarded  as  somewhat  abnormal  pellagra  symptoms.  The 
first  symptoms  appeared  on  the  scrotum,  which  occurs  sometimes, 
as  we  pointed  out  previously.  It  is  apparent,  according  to  Gold- 
berger, that  the  11  subjects  had  pellagra  in  a  mild  form,  and  that 
the  symptoms  may  vary  with  the  diet.  Goldberger  (1365)  held 
that  three  months  sufficed  to  develop  symptoms  of  pellagra  on  a 
suitably  chosen  diet. 


366  THE   VITAMINES 

THEEAPY 

Roussel  (I.e.  1279)  stated  in  1866  that  dietetic  measures  were  of 
greatest  importance  in  combating  pellagra,  a  view  in  which  a  great 
number  of  investigators  are  agreed,  including  Royer  (1366)  who 
expressed  himself  similarly  in  1835.  Lombroso  (I.e.  1325)  saw  that 
a  meat  addition  produces  an  extraordinarily  favorable  effect.  Kleim- 
inger  (1367)  cured  12  pellagrous  lunatics  by  means  of  diet,  and 
Lorenz  (1368)  made  considerable  use  of  the  dieto-therapy.  Elebash 
(1369)  used  green  vegetables  and  fruit  juices;  Allison  (1370)  used 
milk,  eggs  and  fruits;  Sylvester  (1371)  at  first  used  fruit  juices,  then 
skim  milk  and  eggs.  Bravetta  (1372),  Willets  (1373)  and  Ridlon 
(1374)  proceeded  similarly.  The  latter  treated  51  cases  with  a  spe- 
cially selected  diet,  48  of  which  recovered.  Goldberger,  Waring  and 
Willets  (1375)  tested,  in  some  asylums,  a  diet  consisting  of  milk,  meat 
and  vegetables,  with  a  decrease  of  carbohydrates,  and  obtained  very 
good  prophylactic  and  therapeutic  results.  Voegtlin,  Neill  and 
Hunter  (1376)  used  a  vitamine  therapy  in  their  practice,  with  the 
following  results;  with  vitamine  B,  no  results  were  evident,  while  with 
vitamine  A,  in  the  form  of  liver  and  thymus  extracts,  some  result  was 
obtained  but  it  was  not  very  marked.  Sherman  (1377)  recommended 
milk  as  a  prophylactic. 

•As  we  have  noted,  the  best  results  were  obtained  by  dietary 
changes;  in  fact,  the  complex  products  designated  as  animal  protein 
proved  effective.  We  have  already  pointed  out  that  with  the  addi- 
tion of  meat,  eggs  or  milk,  not  animal  protein  alone  is  added,  but  also 
food  complexes  which  are,  for  the  most  part,  unknown.  By  this 
means,  a  diet  rich  in  carbohydrates  and  poor  in  protein  is  changed 
to  one  poor  in  carbohydrates  and  rich  in  protein,  whereupon  the 
entire  nutrition  undergoes  a  radical  change.  The  modern  conception 
of  the  etiology  of  pellagra  is  based  upon  the  above  findings. 

ETIOLOGY   OF   PELLAGRA 

Pellagra  occurs  chiefly  on  corn.  In  this  connection,  a  series  of 
hypotheses  were  proposed  to  explain  the  nature  of  pellagra,  which, 
however,  we  can  not  discuss  here.  Even  the  proponents  of  the  infec- 
tion theory,  for  example,  Slier,  Garrison  and  MacNeal  (1378)  partially 
admit  that  the  diet  also  plays  a  role  in  this  disease.  Tanner  and 
Echols  (1378)  point  out  that  it  is  not  enough  merely  to  give  a 


PELLAGRA  367 

satisfactory  diet.  It  must  also  be  noted  that  the  diet  is  actually 
consumed. 

As  regards  the  nutritive  value  of  corn,  we  have  already  discussed 
this  matter  in  detail  (p.  250).  With  the  assumption  that  even  the 
whole  kernel  possesses  a  low  biological  value,  we  are  not  in  accord, 
since  most  birds  can  live  on  it  indefinitely.  What  happens  to  the 
nutritive  value  of  the  protein,  after  certain  parts  of  the  grain  are 
removed  by  milling,  we  do  not  know  exactly.  The  reports  from 
Egypt  by  Wilson  (I.e.  1330),  Boyd  (1379)  and  Roaf  (1380)  have  shown 
that  in  pellagra,  the  protein  assimilation  is  disturbed,  with  simulta- 
neous defects  of  the  sympathetic  nervous  system.  Wilson,  in  par- 
ticular, is  of  the  opinion  that  the  biological  value  of  the  protein  can 
not  drop  below  40  grams  casein,  without  the  danger  of  the  occurrence 
of  pellagra.  As  a  matter  of  fact,  the  pellagra-producing  diet  there 
was  equal  to  only  22  grams  of  casein. 

Most  of  the  observers  of  the  Egyptian  pellagra  came  to  the  con- 
clusion that  it  is  related  to  a  lack  of  an  essential  aminoacid,  probably 
tryptophane.  Bigland  (1381)  adopted  this  view,  while  Enright 
(1382)  saw  pellagra  in  German  prisoners  who,  in  his  opinion,  had 
sufficient  protein  in  the  diet.  Based  upon  Wilson's  conception, 
Chick  and  Hume  (I.e.  460)  undertook  to  produce  experimental 
pellagra  in  monkeys.  These  monkeys  were  given  a  diet  containing  all 
the  vitamines,  but  varying  considerably  in  its  protein  content  in  the 
form  of  maize  gluten.  Three  animals  were  kept  on  this  diet,  where- 
upon an  erythema  on  the  nostrils  and  other  symptoms  developed 
which  were  regarded  as  pellagra,  and  which  were  improved  by  the 
administration  of  casein.  Since  in  this  experiment,  the  food  and 
vitamine  intake  were  not  controlled,  and  since  the  study  of  a  disease 
supposedly  related  to  a  lack  of  animal  protein  was,  strangely  enough, 
conducted  with  plant-eating  animals,  which  normally  after  the 
suckling  period  never  eat  animal  protein,  we  must  discount  the  value 
of  this  experiment. 

The  conception  that  pellagra  is  associated  with  a  lack  of  protein 
of  high  biological  value,  is  in  some  respects  in  opposition  to  Hind- 
hede's  findings  and  the  experience  of  specialists  connected  with  the 
feeding  of  the  populations  of  various  countries.  In  Europe  too, 
during  the  World  War,  very  little  pellagra  prevailed  despite  the 
lack  of  animal  protein. 


368  THE   VITAMINES 

Infantile  pellagra,  as  for  example  in  the  case  described  by  Voegtlin 
and  Harries  (I.e.  1304),  might  be  of  help  in  clearing  up  the  etiology. 
It  is  indeed  true,  according  to  Eckles,  Palmer  and  Swett  (1383), 
that  the  protein  content  of  the  milk  can  be  influenced  by  nutrition 
to  a  certain  extent;  still,  the  possible  variations  could  hardly  explain 
the  occurrence  of  infantile  pellagra,  of  which  some  instances  have 
been  observed. 

We  are  inclined  to  the  view  of  Goldberger  (I.e.  109),  who  con- 
sidered the  following  etiological  factors  as  possible: 

1.  Partial  lack  of  vitamines. 

2.  Lack  of  animal  protein. 

3.  Lack  of  a  still  unknown  vitamine. 

4.  The  combined  influence  of  all  these  factors. 

We  believe  that  the  dilution  of  protein  with  carbohydrates  increases 
the  vitamine  requirements,  so  that  although  the  vitamines  are 
present,  they  do  not  suffice.  We  have  mentioned  the  possible 
significance  of  the  important  substance,  associated  with  proteins,  and 
it  is  not  impossible  that  this  factor  is  lost  during  the  milling  of  corn. 
It  would  be  rather  premature  to  consider  the  chapter  of  pellagra  as 
closed. 


CHAPTER  VII 
SPRUE 

This  disease,  also  designated  as  aphthae  tropicae,  Ceylon  sore 
mouth,  psilosis  linguae  et  intestini,  diarrhea  alba,  tropical  diarrhea 
and  diarrhee  de  Cochinchine,  has  been  described  in  detail  by  Van  der 
Sheer  (1384),  Thin  (1385),  W.  C.  Brown  (1386),  Begg  (1387),  Schilling 
(I.e.  840  and  980)  and  Castellani  and  Chalmers  (I.e.  839).  This  non- 
infectious  disease,  investigated  since  1776,  is  endemic  in  Asia,  espe- 
cially in  the  Malay  Archipelago,  Siam,  Annam,  Sumatra,  Java,  but 
also  in  India,  Ceylon,  China  and  Japan,  also  in  Australia  and  occa- 
sionally in  Europe.  In  the  United  States,  cases  were  reported  by 
Wood  (1388),  Boyd  (1389)  and  Sturtevant  (1390).  These  patients 
were  never  in  the  tropics,  but  Hiatt  and  Allan  (1391)  also  observed 
cases,  which  came  from  the  tropics.  The  symptoms  of  the  disease 
consist  in  painful  stomatitis  with  vesicles  and  erosions  and  persistent 
diarrhea,  with  grayish-white  foamy  stools.  The  disease  is  curable  by 
a  dietary  therapy. 

SYMPTOMATOLOGY 

The  edge  of  the  tongue  and  the  point  are  reddened,  eroded,  with 
small  vesicles,  and  covered  with  ulcers.  Similar  erosions,  vesicles 
and  ulcers  are  found  on  the  gums,  lips,  soft  palate,  uvula,  and 
cheeks.  The  patients  complain  of  marked  pain  on  chewing  and 
swallowing.  According  to  Bahr  (1392),  the  disease  occurs  much 
more  frequently  in  women,  and  may  be  confused  with  pernicious 
anemia. 

The  abdomen  is  'blown  up,  especially  at  the  epigastrium,  the 
patient  complains  of  pressure,  fullness  and  burning  in  the  region  of 
the  stomach,  particularly  after  meals;  in  addition,  there  is  heart-burn, 
flatulency  and  vomiting  (without  nausea).  On  investigation  of  the 
stomach,  hyperchlorhydria  and  achlorhydria  are  not  infrequently 
found.  In  the  morning  hours,  without  bodily  pain  and  without 
tenesmus,  there  is  a  copious,  soft,  foamy  grayish  evacuation.  The  fat 
content  of  the  feces  is  markedly  increased,  and  the  findings,  according 
to  Halberkann  (1393)  ^are  similar  to  those  in  a  pancreas  affection. 
According  to  T.  B.  Brown  (1393a),  there  is  a  complete  lack  of 

369 


370  THE   VITAMINES 

pancreatic  ferments  and  the  therapeutic  administration  of  such  is 
recommended.  The  number  of  red  blood  corpuscles  is  considerably 
decreased  (1,000,000  to  3,000,000  per  cubic  millimeter);  the  hemo- 
globin index  is  60  to  70  per  cent.  In  some  cases,  tetany  belongs  to 
the  clinical  picture  of  sprue.  In  this  connection,  there  are  the  newer 
reports  of  Bassett-Smith  (1394)  and  Barach  and  Murray  (1395),  who 
investigated  the  parathyroids  and  found  them  normal. 

In  severe  cases,  muscle  weakness,  loss  of  weight,  depression,  and 
symptoms  of  cerebral  irritation  are  observed.  In  the  final  and  fatal 
stage,  continuous  fever  is  often  noted.  The  skin  becomes  gray  and 
scaly;  the  tongue  appears  smooth,  rimous  and  atrophic;  the  ankles 
become  edematous;  the  liver  becomes  atrophic;  the  diarrhea,  more 
severe  and  the  pulse,  slow  and  weak. 

PATHOLOGICAL   ANATOMY 

In  the  intestine,  there  is  a  primary  congestion  of  the  capillaries  of 
the  sub-mucosa;  later,  hemoglobin  exudation  and  round  cell  infiltra- 
tion; finally,  atrophy  of  the  intestinal  mucosa.  This  constant  finding 
of  primary  lesions  in  the  sub-mucosa  speaks  against  a  local  infectious 
cause  of  the  disease.  The  oesophagus  and  the  stomach  are  similarly 
affected,  but  less  intensively.  In  severe  chronic  cases,  atrophy  of 
the  intestinal  wall,  liver  and  pancreas  is  found. 

THEKAPY 

Substances  like  opium,  bismuth,  etc.,  are  rather  harmful.  Emetin 
was  recommended  by  Schmitter  (1396)  although,  according  to 
Miihlens  (1397),  this  is  indicated  only  in  cases  complicated  by 
dysentery.  Simon  (1398)  used  vaccine  with  good  results,  although 
he  stated  that  a  change  in  diet  must  be  made  simultaneously. 
Castellani  (1398a)  recommends  sodium  bicarbonate  administration 
in  conjunction  with  the  dietetic  treatment.  According  to  Wegele 
(1399)  the  dietetic  therapy  is  most  productive  of  results.  Among 
the  best  substances  to  use  are,  according  to  Low  (1400),  milk; 
according  to  Cantlie  (1401),  fruit,  and  according  to  Conran  (1402), 
meat.  These  are  the  same  substances  used  in  the  treatment  of 
avitaminoses,  but  in  sprue,  special  care  must  be  exercised  and  only 
frequent  small  doses  should  be  administered.  Especially  to  be 
recommended,  is  milk,  either  raw  or  cooked  for  a  short  time,  raw 


SPRUE  371 

meat  juice,  meat  broth  and  meat  jelly;  of  the  fruits,  especially  straw- 
berries (2  to  3  pounds  daily),  but  also  apples,  oranges/ grapes,  goose- 
berries, and  blackberries.  Cantlie  (1403)  advised  beginning  carefully 
with  the  meat  diet  (meat  juice,  meat  jelly)  and  giving  milk  every  3 
or  4  days.  An  exclusive  milk  diet  does  not  suffice  to  bring  about  a 
cure,  so  that  in  certain  cases  it  must  be  combined  with  a  fruit  diet. 
Later,  light  mixed  food  is  gradually  and  carefully  given.  Bovaird 
(1403a)  suggests  as  therapy  the  replacing  of  fats  and  carbohydrates 
by  proteins  and  fruits. 

PATHOGENESIS 

Since  sprue  occurs  less  frequently  than  pellagra  the  number  of 
published  communications  on  the  subject  is  not  very  large.  Al- 
though some  investigators  like  Cantlie  (I.e.  1401)  and  Michael  (1404) 
believe  in  an  infectious  cause  (a  fungus,  Monilia  psilosis),  neverthe- 
less the  following  circumstances  point  to  an  avitaminosis :  Ashf ord 
found  (1405)  that  in  Porto  Rico  only  the  city  dwellers,  who  eat 
much  bread,  develop  sprue,  but  not  the  farmers,  who  eat  bananas. 
According  to  W.  C.  Brown  (I.e.  1386),  the  Chinese  coolies  in  East 
India  develop  beriberi,  whereas  the  Europeans,  on  a  practically  similar 
diet,  develop  sprue.  Stewart  (1406)  is  of  the  opinion  that  no  real 
basis  exists  for  regarding  pellagra  and  sprue  as  two  different  diseases. 
Heat  on  (1406a)  believes  rather  in  the  dietetic  than  in  the  infectious 
origin  of  sprue.  Werner  (1407)  observed  scorbutic  symptoms 
(petechiae  on  the  calves  of  the  legs)  in  sprue.  In  the  early  stage  of 
sprue,  there  is  often  an  aversion  for  meat  and  this  feeling,  together 
with  the  known  fear  of  fruit  in  diarrhea,  leads  to  the  further  develop- 
ment of  the  disease.  Still,  the  most  important  reason  for  considering 
sprue  here  is  the  influence  of  dietary  changes  on  the  course  of  the 
disease,  and  the  therapeutic  results  following  the  administration  of 
vitamine-containing  foodstuffs.  In  particular,  the  fact  that  emphasis 
is  placed  upon  the  raw  state  of  the  foodstuffs  in  the  treatment  gives 
further  support  to  our  conception.  Especially  in  the  use  of  straw- 
berries, does  Leede  (1408)  emphatically  state  that  preserved  straw- 
berries are  far  less  effective  than  fresh  berries,  and  that  the  active 
substance  contained  therein  is  destroyed  on  heating.  Whether  we 
have,  in  sprue,  a  disease  similar  to  pellagra  or  scurvy,  cannot  be 
stated  with  certainty  at  present;  we  must  await  further  work  on 
the  subject. 


CHAPTER  VIII 

HUNGER  EDEMA 

Hunger  edema  had  been  observed,  prior  to  the  late  war,  and 
Sticker  (1409)  and  Prinzing  (1410)  occupied  themselves  particularly 
with  the  history  of  this  disease.  Prinzing,  in  his  historical  compi- 
lation, records  no  cases  between  the  time  of  the  Peloponesian  War 
(430  to  425  B.C.)  and  the  siege  of  Port  Arthur  in  1904.  Maliwa 
(1411)  believed  that  hunger  edema  was  observed  in  the  Napoleonic 
Wars,  at  the  siege  of  Paris,  and  in  the  Boer  War.  Wheeler  (1412) 
described  actual  cases  in  the  Boer  War,  in  the  concentration  camps. 
Digby  (1413)  and  McLeod  (1413a)  reported  on  this  disease  during 
famines  in  India  and  Ceylon  in  1876  and  1877.  Patterson  (1414) 
saw  numerous  cases  in  Chinkiang  (China)  in  1899,  during  the  time 
that  the  population  was  living  on  greens.  Landa  (1415)  counted 
hundreds  of  cases  in  Mexico  during  the  war  in  1915,  which  developed 
on  a  diet  of  turnips  and  spinach. 

In  the  last  war,  the  first  report  on  the  subject  was  made  by  Rumpel 
(1416).  Then,  in  rapid  succession  there  came  numerous  reports  of 
the  disease  among  prisoners  by  Jtirgens  (1417),  Bonheim  (1418)  in 
Bonn  and  Lange  (1419)  in  East  Prussia.  From  Austria,  among 
other  reports,  were  those  of  Knack  (1420),  Schiff  (1421)  and  Jaksche 
(1422) ;  indeed,  in  one  part  of  Bohemia  alone,  22,000  cases  were  noted, 
with  4  per  cent  mortality. 

Vandervelde  and  Cantineau  (1423)  and  Breuer  (1424)  reported  on 
cases  in  Belgium.  Beyerman  (1425)  observed  cases  in  the  insane 
asylum  in  Medemblik  (Holland)  in  1917  and  1918,  which  were  cured 
by  fresh  vegetables;  in  these  cases  there  was  perhaps  a  complication 
with  scurvy.  Strauss  (1426),  Guillemin  and  Guyot  (1427)  and,  at 
the  beginning  of  the  war,  Budzynski  and  Chelchowski  (1428)  de- 
scribed the  disease  in  Poland.  Wells  (1429)  saw  numerous  cases 
in  1917  in  Roumania;  Tonin  (1430)  observed  the  disease  among 
prisoners  in  Italy;  Enright  (1431),  among  Turkish  prisoners  in 
Egypt. 

Mann,  Helm  and  Brown  (1432)  noted  3000  cases  in  Haiti,  out  of 
which  200  came  to  necropsy.  The  disease  has  prevailed  there  for 

372 


HUNGER  EDEMA  373 

years  among  the  prisoners,  in  fact,  it  was  noted  that  the  disease 
broke  out  in  less  than  three  months  after  incarceration. 

In  infants,  on  a  carbohydrate-rich  diet,  there  develops  a  condition 
called  "Mehlnahrschaden."  of  which  we  have  already  spoken. 
Many  of  these  cases,  if  not  all,  can  be  considered  analogous  to  hunger 
edema.  Vacher  (1433)  reported  such  cases  during  the  siege  of  Paris; 
later,  similar  conditions  were  described  by  de  Wolf  (1434)  and 
Potter  (1435)  in  children  suffering  from  marasmus.  Potter  believed 
that  lack  of  protein  was  the  cause  of  the  disease.  The  disease  was 
also  noted  by  Chapin  (1436),  Waterman  (1437),  Hume  (1438),  Ashby 
(1439)  and  Klose  (1440).  Many  of  these  investigators  studied  the 
effect  of  a  change  in  the  diet,  especially  an  addition  of  protein. 
In  the  American  literature,  the  disease  is  often  designated  as  acro- 
dynia  and  has  been  described  by  Weston  (1440a)  and  by  Cartin 
(1440b). 

Of  additional  interest,  is  the  relationship  between  hunger  edema 
and  "epidemic  dropsy"  in  India.  This  disease,  described  by 
Greig  (I.e.  880),  was  first  regarded  as  wet  beriberi.  Although 
scorbutic  symptoms  were  also  observed,  the  condition  of  the  patients 
could  be  markedly  improved  by  an  addition  of  meat.  Most  of  these 
cases  developed  on  a  rice  diet,  and  yet  they  could  be  identified  as 
hunger  edema.  McCay  (1441)  believed  that  the  cause  of  epidemic 
dropsy  lay  in  the  lack  of  protein.  In  a  compilation  on  this  subject, 
Maver  (1442)  believed  that  hunger  edema,  edema  of  children  and 
epidemic  dropsy  have  the  same  etiology. 

SYMPTOMATOLOGY  AND  MODE  OF  DEVELOPMENT 

A  splendid  description  of  the  symptomatology,  pathology  and 
pathogenesis  is  given  by  Schittenhelm  and  Schlecht  (1443)  and  by 
Schiff  (1444).  The  most  prominent  sign  is  the  edema,  localized 
mostly  on  the  legs  and  on  the  face,  near  the  eyes.  According  to 
Htilse  (1445)  pains  in  the  calves  are  very  characteristic.  Polyuria 
is  very  pronounced,  according  to  Rumpel  and  Knack  (1446)  and 
Zondek  (1447).  Jansen  (1448)  found  decreased  blood  pressure  and 
subnormal  temperatures.  Jess  (1449)  called  attention  to  the  eye 
symptoms,  hemeralopia  and  peripapillary  edema  of  the  retina.  The 
marked  bradycardia  is  especially  characteristic  of  hunger  edema, 
according  to  Rumpel  and  Knack  (I.e.  1446).  This  is  not  to  be  at- 


374  THE   VITAMINES 

tributed  to  heart  weakness,  according  to  Schittenhelm  and  Schlecht 
(I.e.  1443),  but  to  the  same  causes  that  bring  about  the  edema.  In 
these  cases,  adrenaline  dose  not  raise  the  blood  pressure,  and  the  usual 
heart  stimulants,  such  as  digitalis  and  caffeine,  have  little  effect, 
while  atropine  has  no  effect  on  the  bradycardia.  The  pulse  is 
extraordinarily  small,  slow  (32  to  36)  and  soft;  reflexes  and  normal; 
constipation  was  noted  more  frequently  than  diarrhea,  though  the 
latter  progressed  as  the  edema  disappeared.  The  disease  appeared 
mostly  in  hard-working  men;  the  over-exertion  seemed  to  hasten 
the  onset  of  the  disease.  Frequently,  rest  in  bed  sufficed  to  dissi- 
pate the  edema  and  to  bring  about  an  improvement.  The  disease 
is  often  found  associated  with  various  infections,  such  as  lobar 
pneumonia,  bronchitis  and  furunculosis. 

Various  investigators  offer  different  explanations  of  the  mode  of 
development  of  the  disease,  though  there  is  no  doubt  that  the  poor 
diet  must  be  the  primary  cause ;  the  predisposing  conditions  may  be 
severe  manual  labor  and  exposure.  Knack  and  Neumann  (1450) 
stated  that  the  cause  was  inadequate  nutrition  with  turnips  and  a 
large  water  intake.  Falta  (1451)  thought  that  the  faulty  diet  was 
bread,  cabbage,  dried  vegetables  and  turnips.  The  total  diet  con- 
tained 30  to  59  grams  protein  and  1200  to  1400  calories;  the  vege- 
tables were  cooked  in  large  quantities  for  4  to  7  hours.  The  cases 
reported  by  Falta  occurred  among  war  prisoners.  Burger  (1452) 
gave  the  exact  composition  of  the  faulty  diet;  it  consisted  of  55.9 
grams  of  protein,  8.5  grams  fat  and  284.6  grams  carbohydrate  (1478 
calories).  Kraus  (1453)  observed  cases  developed  after  hard  labor, 
on  a  diet  containing  15  per  cent  indigestible  carbohydrates,  little 
fat  and  a  maximum  of  50  grams  protein,  and  800  to  1300  calories. 

PATHOLOGY 

On  autopsy,  a  complete  disappearance  of  subcutaneous  fat  tissue 
was  found,  although  well-nourished  bodies  also  came  to  necropsy. 
Atrophy  of  all  parenchymatous  organs  was  observed;  the  liver  and 
muscles  were  glycogen-free.  Hlilse  (1454)  found  atrophy  of  the 
heart;  in  a  few  cases  the  adrenals  were  enlarged;  however,  according 
to  Lippmann  (1455),  no  pathological  changes  in  the  nerves  were 
observed.  According  to  Paltauf  (1456)  the  anemia  is  very  insignifi- 
cant. In  the  pre-edematous  stage,  according  to  Jacobsthal  (1457) 


HUNGER   EDEMA  375 

high  hemoglobin  and  erythrocyte  values  were  found.  Woltmann 
(1458)  found  anisocytosis,  polychromatophilia,  high  leucopenia, 
lymphocytosis  and  hydration  of  the  blood.  In  epidemic  dropsy, 
Maynard  (1459)  found  increased  intra-ocular  pressure. 

METABOLISM 

Schittenhelm  and  Schlecht  (I.e.  1443)  conducted  metabolism 
experiments  with  a  diet  regarded  as  productive  of  edema  (45  to  55 
grams  protein  and  1100  to  1800  calories).  It  was  soon  established 
that  a  characteristic  assimilative  failure  was  not  shown  by  the 
patients,  since  they  gained  weight  on  more  protein  and  fat  and  re- 
tained nitrogen.  Zondek  (I.e.  1447)  had  some  patients  who  excreted 
six  liters  of  urine  daily,  with  40  grams,  and  more,  of  sodium  chlo- 
ride, accompanied  by  a  normal  nitrogen  content.  Phosphoric  acid 
excretion  was  high  (4  to  5  grams);  Hiilse  (I.e.  1454)  was  able  to 
demonstrate  a  good  protein  assimilation,  but  believed  that  the 
de-aminization  of  the  amino  acids  was  faulty.  Franke  and  Gottes- 
mann  (1460)  studied  the  rate  of  elimination  of  urea,  sodium 
chloride  and  other  salts,  and  found  a  retardation.  The  pathological 
chemistry  of  hunger  edema  was  considered  by  Feigl  (1461),  who 
studied  the  cases  of  Rumpel.  Schittenhelm  and  Schlecht  (I.e.  1443) 
found  the  protein  content  of  the  blood  low,  4.5  to  5.7  per  cent  instead 
of  7  to  9  per  cent.  The  erythrocytes  showed  a  lowered  lipoid  con- 
tent. In  the  opinion  of  these  investigators,  the  low  figures  are  not 
to  be  attributed  merely  to  the  dilution  of  the  blood,  for  as  the  edema 
disappears,  the  blood  values  drop  still  more.  The  blood  sugar  is 
either  normal  or  increased.  The  composition  of  the  edematous 
fluid  resembles  that  of  exudates.  At  the  height  of  the  edema,  a 
retention  of  water  and  chlorides  is  noted.  On  the  disappearance  of 
the  edema,  considerable  urea  and  sodium  chloride  excretion  is  found, 
which  is  increased  still  more  by  thyroidin  and  decreased  by  adrenaline. 
Pituitary  extracts  are  effective,  but  only  on  the  sodium  chloride 
excretion. 

THERAPY 

Here  too,  there  is  no  uniformity  in  the  literature.  Zak  (1462) 
believed  that  hunger  edema  could  be  cured  not  only  through  the 
addition  of  animal  products  (eggs,  raw  calves'  liver),  but  also  by 
means  of  carrot  juice.  Maase  and  Zondek  (1463)  were  of  the  opinion 
that  the  addition  of  100  grams  fat  could  arrest  the  progress  of  the 


376  THE   VITAMINES 

disease.  Schiff  (I.e.  1444)  sought  the  cause  of  the  disease  not  in 
the  lack  of  calories,  per  se,  but  of  potatoes,  fresh  vegetables  and 
milk;  addition  of  yeast,  beer  and  fat  had  no  therapeutic  effect. 
Knack  and  Neumann  (I.e.  1450)  obtained  good  results  with  potatoes. 
Reach  (1464)  made  use  of  organotherapy,  in  the  form  of  testicles, 
with  good  results,  v.  Hosslin  (1465)  and  Schittenhelm  (1466) 
believed  mainly  in  the  effect  of  protein  therapy.  Isenschmid  (1467) 
attributed  certain  edemas,  like  those  occurring  after  dysentery  and 
other  protracted  diseases,  to  the  same  causes  that  bring  about 
hunger  edema,  and  in  his  opinion,  they  should  be  treated  in  a  similar 
manner.  Most  investigators  manifest  a  particular  belief  in  protein 
of  animal  origin  as  a  therapeutic  measure.  The  above  therapy  con- 
stitutes the  sum  and  substance  of  the  leading  ideas  as  regards  the 
etiology  of  hunger  edema. 

ETIOLOGY 

Because  of  the  great  lack  of  fat  during  the  war,  some  investigators 
associated  the  etiology  with  this  factor.  We  have  just  seen  that 
this  is  not  likely  and  that  it  has  to  do  chiefly  with  a  lack  of  vitamine 
A,  and  most  animal  products  used  contain  more  or  less  of  this  sub- 
stance. Aron  (1467a)  considers  that  hunger  edema  is  due  to  an 
insufficient  amount  of  calories  in  the  diet.  Park  (1468)  was  of  the 
opinion  that  the  edema  was  associated  with  a  lack  of  protein  and  fat, 
and  an  excessive  water  intake.  Rubner  (1469)  and  Determann  (1470) 
attributed  it  to  the  lack  of  protein.  The  latter  conception  is  also  in 
accord  with  the  experience  gained  in  the  studies  on  parenchymatous 
nephritis  by  Epstein  (1471)  and  on  renal  edema  by  Allbutt  (1472). 
Further  confirmation  is  given  by  the  study  of  the  development  of 
experimental  edema  in  animals,  especially  of  rats  and  mice.  Tachau 
(I.e.  1244)  stated  that  when  the  correct  relationship  between  proteins 
and  carbohydrates  is  disturbed  by  the  addition  of  the  latter,  edema 
develops.  Denton  and  Kohman  (1473)  observed  that  when  rats 
are  fed  on  carrots,  casein,  starch,  butter  or  lard  and  salts,  this  diet 
is  adequate  only  when  the  carrots  provide  half  of  the  total  calories. 
However,  if  this  relationship  is  changed  by  the  addition  of  starch  or 
fat,  then  edema  develops  in  a  great  percentage  of  the  animals. 
Kohman  (1474)  carried  this  idea  further,  in  that  by  means  of  the 
above  diet,  especially  with  the  addition  of  water,  the  animals 
developed  edema,  which  was  cured  by  the  administration  of  casein. 
No  exact  figures  are  given  as  to  the  food  intake,  but  in  any  event, 


HUNGER   EDEMA  377 

we  have  already  voiced  our  opinion,  in  the  introduction  to  this  chap- 
ter, against  the  conception  of  a  lack  of  protein  as  an  etiological 
factor  in  the  development  of  hunger  edema.  By  the  addition  of 
casein,  it  is  possible,  first  of  all,  that  there  might  have  been  an 
increase  in  appetite,  with  a  consequently  greater  food  and  vitamine 
intake.  The  considerable  quantity  of  water  not  only  diluted  the 
vitamine  and  protein  content  of  this  diet,  but  also  caused  great  loss 
by  washing  out.  Kohman  tried  to  get  around  these  difficulties  by 
showing  that  on  a  synthetic  diet  prepared  in  the  usual  manner,  a 
small  food  intake,  per  se.  is  not  followed  by  edema.  However,  since 
the  diet  was  properly  constituted,  in  this  case,  the  results  cannot 
be  applied  to  the  carrot  diet. 

Another  hypothesis  deals  with  the  role  of  the  adrenals  in  the 
development  of  edemas;  this  was  formulated  by  McCarrison  (I.e. 
290  and  608).  He  showed  that  in  beriberi,  there  was  an  increased 
adrenaline  output  which  was  supposed  to  be  responsible  for  the 
development  of  edema.  McCarrison  held  that  butter,  but  not  all 
fats,  contains  a  substance  (apparently  vitamine  A)  which  is  protec- 
tive against  edema.  This  idea  was  adopted  by  Bigland  (1475),  in 
a  practically  unchanged  form,  who  regarded  an  increased  or  de- 
creased suprarenal  activity  as  the  causative  factor  in  the  explanation 
of  hunger  edema  and  pellagra,  respectively.  Nixon  (1476)  assumed, 
as  the  cause  of  edema,  insufficient  calories  (fat?)  and  protein  and 
excess  of  water;  adrenaline  was  also  thought  to  play  a  part.  In 
conclusion,  we  shall  mention  the  pathogenesis  assumed  by  Schitten- 
helm  and  Schlecht  (I.e.  1443).  They  ascribed  the  cause  to  the 
lack  of  protein  and  fat,  the  effect  of  cold  and  hard  labor  helping  to 
bring  about  the  disease.  They  recognized  that  hunger  edema  occurs 
on  a  diet  rich  in  carbohydrates.  The  differentiation  from  beriberi 
lies  in  the  bradycardia;  on  the  other  hand,  scurvy-like  symptoms, 
such  as  stomatitis  and  bleeding  of  the  gums  were  observed. 
Hemeralopia  and  xerosis  of  the  eyes  were  also  noted  quite  frequently. 
Administration  of  vitamine  B  exerted  no  therapeutic  effect.  There 
is  a  great  analogy  to  "Mehlnahrschaden." 

Our  presumption  is  that  we  are  dealing  here  with  a  complication 
of  causes,  although  the  disproportion  between  the  individual  dietary 
constituents,  as  well  as  an  inadequate  diet  in  relationship  with  the 
hard  labor  stands  in  the  foreground.  However,  in  this  case  too, 
further  work  is  necessary  in  order  to  arrive  at  an  explanation  of  the 
problem. 


CHAPTER  IX 

PATHOLOGICAL  CONDITIONS  IN  WHICH  THE  LACK  OF  VITAMINES 
MAY  BE  SUSPECTED 

It  is  difficult,  if  not  impossible,  to  determine  in  the  conditions 
mentioned  below,  at  the  present  time,  if  the  described  effect  is  due 
to  the  vitamines,  per  se,  or  to  the  other  nutritive  constituents  admin- 
istered simultaneously.  The  subject  may  be  cleared  up  only  when 
we  shall  have  the  vitamines  available  in  larger  amounts  in  the  pure  or 
almost  pure  state. 

KALLAK 

Little  (1477)  reported,  in  1908,  on  a  disease  called  "kallak," 
occurring  among  the  Eskimos  in  New  Foundland.  The  disease 
consisted  in  a  marked  pustulous  dermatitis  on  the  hands,  elbows 
and  other  parts  of  the  body,  with  persistent  itching.  The  disease 
is  usually  curable.  The  natives  already  know,  from  experience, 
that  the  disease  makes  its  appearance  when  only  a  small  quantity 
of  seal  meat  is  available.  It  occurs  in  autumn,  when  they  live 
mostly  on  fish  and  when  wild  berries  become  scarce.  Their  diet 
consists  of  seal  meat,  carribou,  birds,  fish  and  berries,  and  is  very 
rich  in  protein.  The  disease  is  very  predisposing  to  scurvy,  and 
Little  believed  that  it  might  be  an  avitaminosis. 

TRENCH   SICKNESS 

Bruntz  and  Spillmann  (1478)  observed  the  disease  among  the 
French  soldiers  in  the  trenches.  It  consists  partly  of  neuritic 
symptoms  (parasthesias,  pains,  motor  disturbances,  asthenia),  and 
scorbutic  symptoms  in  the  mouth,  especially  the  gums.  Following 
a  better  diet  (fresh  vegetables  and  fruits),  the  number  of  cases 
decreased  markedly.  It  is  related  to  scurvy,  with  a  partial  lack  of 
other  vitamines,  and  is  made  more  severe  by  cold  and  over-exertion. 
Psychic  phenomena  and  uncleanliness  apparently  also  play  a  great 
role  in  the  pathogenesis.  All  the  extremities  were  affected.  This 
disease  was  also  studied  by  Mercier  (1479)  and  Chauvin  (1480). 

378 


PATHOLOGICAL    CONDITIONS   DUE   TO   LACK   OF   VITAMINES        379 
INTESTINAL   STASIS,    ETC. 

Following  his  interesting  findings  in  the  intestines  in  avian  beriberi, 
McCarrison  (1481)  sought  to  apply  his  results  to  an  explanation 
of  some  obscure  intestinal  disorders.  He  thought  for  example,  that 
properly  nourished  children  showed  less  tendency  towards  intussus- 
ception. Among  the  diseases  which  McCarrison  associated  with 
the  vitamines  is  intestinal  stasis,  which  was  treated  by  Sir  Arbuthnot 
Lane  (1482),  and  later,  intestinal  toxemia  and  the  so-called  "coeliac 
disease,"  described  by  Still  (1483).  The  latter  observed,  among 
41  cases,  4  that  had  scurvy  previous  to  developing  coeliac  disease. 
As  an  indication  of  the  lack  of  vitamines,  McCarrison  pointed  out 
dilatation  of  the  stomach,  air  pockets  in  the  intestine,  impairment 
of  the  neuro-vascular  control  of  the  gastro-intestinal  tract,  formation 
of  stomach  ulcer  and  especially  colitis.  Mackenzie  Wallis  (1484) 
adopted  the  view  of  McCarrison  in  regard  to  coeliac  disease,  in  that 
he  made  use  of  vitamine  therapy  in  this  condition  among  children, 
with  good  results. 

STERILITY 

Reynolds  and  Macomber  (1485)  found  that  when  rats  are  fed 
on  inadequate  diets  (also  in  the  case  of  vitamine  deficiency)  they 
exhibit  a  marked  diminution  in  fertility.  This  viewpoint  which  is 
well  borne  out  by  laboratory  experience  has  been  considered  in  the 
treatment  of  analogous  conditions  in  man  and  favorable  results  have 
been  reported. 

HEMERALOPIA 

We  have  already  seen  that  in  the  various  human  avitaminoses, 
hemeralopia  was  frequently  observed.  This  is  not  associated  with 
any  particular  vitamine,  but  arises  unspecifically,  perhaps  on  any 
kind  of  inadequate  diet.  We  have  nevertheless  found  data  in  the 
literature  pointing  to  a  widespread  occurrence  of  this  eye  affection, 
which  was  favorably  influenced  by  cod  liver  oil.  Ishiwara  (1486) 
saw  such  cases  in  Japan.  He  studied  the  lipoid  content  of  the  blood 
and  found  it  decreased.  When  the  disease  was  cured,  the  figures 
returned  to  normal.  Tricoire  (1487)  observed  among  French  soldiers, 
320  such  cases  which  were  promptly  cured  by  cod  liver  oil. 
H.  Smith  (1487a)  is  of  the  opinion  that  hemeralopia  has  mostly  been 
simulated  during  the  war  since  the  visual  field  was  not  diminished  as 


380  THE   VITAMINES 

in  true  cases.  McCollum,  Simmonds  and  Parsons  (I.e.  1222a)  re- 
gard hemeralopia  as  of  dietetic  origin,  and  curable  with  cod  liver 
oil.  Appleton  (1487b)  considers  hemeralopia  and  snow  blindness, 
as  it  occurs  in  polar  regions  (Labrador),  due  to  the  strong  sun  rays 
on  the  basis  of  a  nervous  disturbance  and  supposed  to  occur  only 
in  men  who  work  in  the  open.  Pick  (1488)  and  Feilchenfeld  (1489) 
reported  on  the  influence  of  war  diets  upon  various  eye  affections. 

EXOPHTHALMIC    GOITRE    (GRAVES   DISEASE) 

In  the  study  of  experimental  rickets  in  dogs,  Edward  and  May 
Mellanby  (I.e.  408a  and  b)  made  the  interesting  observation  that 
on  feeding  certain  fats,  like  butter,  a  hyperplasia  of  the  thyroid 
develops  which  can  be  cured  by  cod  liver  oil.  This  observation  has 
been  made  use  of  in  human  cases  and  good  results  have  been  obtained, 
though  the  number  of  cases  up  to  the  present  have  been  insufficient 
to  permit  of  definite  conclusions  being  made. 

THE    SIGNIFICANCE    OF   VITAMINES   IN   INFECTIONS 

Since  an  inadequate  diet,  especially  the  deficiency  in  vitamines, 
lowers  the  resistance  towards  infections,  an  insufficient  production 
of  antibodies  was  first  thought  of  as  an  explanation.  The  question 
has  been  studied  experimentally  by  some  investigators.  Hektoen 
(1490),  in  rats  on  artificial  diet,  and  Zilva  (1491),  in  rats  and  guinea 
pigs  on  a  vitamine-poor  diet,  observed  a  normal  formation  of  anti- 
bodies. On  the  other  hand,  according  to  Kleinschmidt  (1492)  the 
hemolysin  formation  does  not  attain  its  normal  value  on  an  un- 
suitable diet.  It  is  well  known  that  the  method  of  feeding  plays  an 
important  part  in  various  infections,  as  has  been  shown  by  E.  Thomas 
(1493),  Valagussa  (1494)  and  Peiser  (1495).  The  latter  reported 
especially  on  the  influence  of  the  dietary  fat  on  the  resistance  to 
infection.  During  the  course  of  our  discussion,  we  have  already 
pointed  out  the  possible  relationship  between  the  vitamines  and 
some  infections,  and  we  must  call  attention  to  the  fact  that  since  the 
recognition  of  the  nature  of  ophthalmia,  this  matter  has  assumed 
greater  significance.  The  known  data  on  ophthalmia  indicate 
perhaps,  that  they  may  be  partially  applied  to  other  infectious 
conditions.  For  here,  we  have  a  local  infection  that  may  be 
influenced  specifically  by  a  vitamine  therapy. 


PATHOLOGICAL    CONDITIONS   DUE   TO   LACK   OF   VITAMINES       381 

A  further  field  in  which  the  vitamines  may  be  of  significance,  is 
the  diet  in  various  acute,  and  especially  chronic  infections.  During 
convalescence,  after  acute  infections,  the  diet  is  of  greatest  impor- 
tance. It  happens  occasionally  that  in  all  these  conditions,  quite 
a  one-sided  diet  is  given  for  weeks  at  a  time,  for  example,  flour 
preparations,  because  of  which  recovery  is  impeded,  and  the  resis- 
tance of  the  patients  is  further  decreased.  These  conditions  occur 
especially  after  typhoid  fever,  as  Morrison  (1496)  showed.  The 
question  of  the  diet  in  typhoid  was  studied  by  Barker  (1497),  while 
Coleman  (1498)  Walton  (1499),  Carter,  Howe  and  Mason  (1500) 
laid  emphasis  on  the  caloric  value  of  the  diet  in  convalescence. 
Coleman  spoke  of  the  necessity  of  a  well-balanced  diet,  with  which 
he  treated  222  patients,  with  the  result  that  the  mortality  was  50 
per  cent  less.  Combe  (1501)  described  the  influence  of  the  war 
diet  on  the  course  and  convalescence  of  typhoid  in  Germany. 

Isenschmid  (I.e.  1447)  believed  that  some  edemas,  which  occur 
after  dysentery  or  other  protracted  diseases,  are  of  dietary  origin  and 
belong  to  the  type  of  hunger  edema.  In  all  these  cases,  it  is  important 
that  the  diet,  even  though  small  in  quantity,  in  accord  with  the 
appetite  of  the  patient,  must  be  complete  in  its  composition.  It  is 
not  without  reason  that  milk  is  so  often  used  for  the  above  purpose. 
If  the  disease  or  convalescence  is  too  long  drawn  out  simultaneously 
with  an  unsuitable  diet,  then  it  is  possible  for  a  real  avitaminosis 
to  develop. 

OPHTHALMIA 

The  disease,  likewise  called  "xerophthalmia,"  seems  to  occur  also 
endemically.  Thus,  Hirdlika  (1502)  described  a  similar  disease 
among  the  Indians  in  the  southwestern  part  of  the  United  States, 
while  McCarrison  (1503)  stated  that  the  disease  occurs  not  infre- 
quently in  India,  on  a  diet  of  rice  and  vegetable  oils,  and  is  curable 
by  cod  liver  oil.  The  disease  consists  in  an  infection  of  the  con- 
junctiva, which,  under  certain  conditions,  may  lead  to  a  destruction 
of  the  cornea,  falling  of  the  lens  and  the  iris,  and  total  blindness. 
Mori  (1504)  was  one  of  the  first  to  describe  the  condition  somewhat 
more  in  detail.  An  outbreak  of  this  disease,  called  "hikan,"  which 
developed  at  a  time  of  partial  famine,  was  mentioned  by  Mori,  who 
investigated  1400  cases  in  detail.  It  rarely  occurred  among  fisher- 
men, and  could  be  cured  with  chicken  liver.  Czerny  and  Keller 


382  THE   VITAMINES 

(1505)  observed  the  development  of  the  disease  on  a  carbohydrate- 
rich  diet.  Falta  and  Noeggerath  (I.e.  18)  were  perhaps  the  first  to 
produce  this  disease  experimentally  in  animals.  Later,  this  con- 
dition was  observed  in  rate  by  Knapp  (1506).  Already  at  that  time, 
he  believed  that  a  similar  condition  could  occur  in  children.  Since 
1913,  there  have  appeared  in  rapid  succession,  the  papers  of  Osborne 
and  Mendel,  as  well  as  those  of  McCollum,  already  mentioned,  the 
cause  of  the  disease  being  associated  with  the  absence  of  certain  fats 
in  the  diet.  In  the  end,  this  disease  was  regarded  as  an  avitaminosis, 
caused  by  the  lack  of  vitamine  A.  This  eye  affection  was  also  studied 
in  Germany  by  Freise,  Goldschmidt  and  Frank  (1507),  working  on 
rats.  Goldschmidt  (1508)  stated  that  the  disease  could  be  cured  by 
a  small  amount  of  skim  milk.  Later  it  was  also  produced  in  mice 
and  rabbits  and  although  Nelson  and  Lamb  (1509)  stated  that  they 
could  not  produce  it  in  guinea  pigs  and  chickens,  we  have  information 
to  the  effect  that  ophthalmia  has  been  produced  in  guinea  pigs. 
We  have  personally  seen  numerous  cases  in  chickens,  terminating  in 
total  blindness.  Recently,  we  have  been  corroborated  by  Guerrero 
and  Conception  (1510),  who  observed  the  disease  in  chickens  fed  on 
white  rice. 

In  recent  years,  ophthalmia  has  been  frequently  noted  in 
children.  A  particularly  careful  investigation  was  made  by  Bloch 
(1511)  in  Copenhagen.  He  observed,  during  a  period  of  5  years  in 
the  children's  clinic,  40  cases  among  children  fed  on  a  highly  centri- 
fuged  milk,  fat  being  given  in  the  form  of  vegetable  margarine.  A 
cure  was  obtained  with  whole  milk  or  cod  liver  oil.  The  diet  in 
these  cases  appeared  to  be  inadequate  also  in  other  respects.  Bloch 
(151  la)  believes  that  ophthalmia  occurs  quite  frequently  in  Den- 
mark and  may  often  be  the  cause  of  blindness.  Monrad  (1512) 
and  R0nne  (1513)  reported  on  the  same  condition  in  Denmark. 
In  addition,  one  case  was  reported  in  France  by  Sztark  (1514)  and 
in  the  United  States  by  Parker  (1515).  Sztark's  case  was  a  27 
months  old  baby  fed  on  vegetable  soups,  without  milk;  cod  liver  oil 
effected  a  cure  in  about  10  days. 

The  nature  of  the  disease 

Lately,  the  investigators  have  been  more  and  more  inclined  to 
assume  that  ophthalmia  is  associated  with  a  lack  of  vitamine  A. 
The  pathogenesis  of  this  disease  is  not  so  very  simple,  since  we  are 


PATHOLOGICAL    CONDITIONS    DUE    TO    LACK    OF   VITAMINES       383 

dealing  here  with  a  new  conception  in  pathology.  According  to 
Macfie  (1516),  it  is  a  non-contagious  infection  which  may  be  pre- 
vented 2nd  cured  by  diet.  Funk  arid  Macallum  (I.e.  86)  reported 
that  they  could  prevent  this  eye  disease  by  antiseptic  treatment,  and 
this  was  concurred  in  by  Bulley  (1517).  Later,  however,  from  the 
same  laboratory  in  which  Bulley  worked,  the  report  of  Stephenson 
and  Clark  (1518)  appeared  refuting  Bulley 's  findings.  They  believed 
that  Bulley's  results  were  due  to  the  contamination  of  the  casein  by 
vitamine  A.  In  these  cases,  the  eyes  were  also  examined  bacteri- 
ologically.  The  infection  was  shown  to  be  unspecific,  and  it  is  of 
interest  that  before  the  development  of  the  disease,  no  histological 
changes  were  found  to  account  for  the  slight  resistance  towards  the 
infection.  In  addition,  Stephenson  and  Clark  found  that  not  all 
animals  lacking  vitamine  A  develop  ophthalmia.  Wason  (1518a), 
who  called  the  disease  " ophthalmia,"  arrived  at  the  same  conclusion 
on  the  basis  of  her  findings.  The  specificity  of  ophthalmia,  as 
being  due  to  lack  of  vitamine  A,  has  recently  been  studied  by  Emmett 
(1519).  His  statistical  data  are  as  follows: 

1.  Without  vitamine  A — 122  rats;  ophthalmia  in  120  =  98.3  per  cent. 

2.  Without  vitamine  B — 103  rats;  ophthalmia  in  0  =  0. 

3.  Normal  diet — 216  rats;  ophthalmia  in  0  =  0. 

This  disease  could  not  be  transmitted  from  one  animal  to  another. 
Mendel  (1520)  observed  no  ophthalmia  among  7000  rats  fed  on 
insufficient  protein,  and  in  225  rats  which  were  given  no  B-vitamine. 
However,  out  of  136  rats  receiving  no  A-vitamine,  69  developed  the 
disease.  The  author  has  personally  never  worked  with  great  num- 
bers of  rats;  we  have  had  usually  about  30  rats  at  one  time  under 
observation,  but  on  several  occasions  we  have  seen  ophthalmia  in 
the  presence  of  sufficient  butter  or  cod  liver  oil.  Thus,  Funk  and 
Dubin  (I.e.  331)  have  recently  noted  two  such  cases  out  of  30  rats. 
These  two  manifested  intermittent  ophthalmia,  lasting  for  a  short 
time,  disappearing,  and  re-appearing  in  a  few  days,  but  permanent 
lesions  were  recognizable.  This  form  was  favorably  influenced  by 
yeast.  It  is  worthy  of  note  that  Guiral  (1521)  reported  ophthalmia 
in  children,  who  were  completely  cured  by  orange  juice  in  8  to  10 
days.  We  are  in  possession  of  the  private  data  of  specialists  who 
observed  the  eye  condition  in  the  presence  of  vitamine  A,  which 
was  favorably  affected  by  orange  juice.  Still,  it  must  certainly  be 


384  THE   VITAMINES 

admitted  that  the  absence  of  vitamine  A  favors  the  occurrence  of 
ophthalmia  in  quite  a  specific  manner,  while  contrary  findings  require 
further  explanation.  It  is  possible,  for  example,  that  in  the  presence 
of  cod  liver  oil,  but  with  a  small  food  intake  there  might  be  insuffi- 
cient vitamine  A  supplied  to  the  animal.  All  substances  containing 
vitamine  A  may  be  used  in  the  therapy  of  ophthalmia. 

TUBERCULOSIS 

That  diet  plays  an  important  part  in  the  therapy  of  tuberculosis, 
is  too  well  known  to  require  further  discussion.  An  added  confirma- 
tion of  this  is  found  in  the  reports  on  the  influence  of  the  war  diets, 
of  which  we  have  already  spoken.  However,  we  may  mention  once 
more  the  communication  of  Adams  and  Hamilton  (I.e.  1048),  dealing 
with  the  increase  in  tuberculosis  in  Germany  during  the  war.  Ham- 
burger (1522)  also,  in  his  report  on  the  war  diet  of  children,  pointed 
out  the  dependence  of  the  number  of  cases  of  tuberculosis  on  the  diet, 
stating  that  it  was  due  to  the  lack  of  fat.  Stolzner  (1523)  likewise 
reported  on  the  effect  of  the  diet.  Geoghegan  (1524)  reported  on 
some  cases  in  the  West  Indies,  which,  in  his  opinion,  are  due  to  the 
prevailing  poverty.  On  these  islands,  with  good  climatic  conditions, 
there  should  be  very  little  tuberculosis.  Kichet  (1525)  believed  that 
meat  juice  contains  ferments  which  act  favorably  on  tuberculosis. 
Woodcock  and  Rustin  (1526)  stated  that  the  diet  in  tuberculosis 
must  be  rich  in  protein  and  fat  (in  the  form  of  milk  or  milk  products) . 
If  margarine  is  used,  it  must  be  oleo-margarine  which  contains  vita- 
mine  A;  the  diet  must  also  contain  plenty  of  fruits  and  vegetables. 
Gar dey  (1526a)  finds  that  certain  symptoms  of  pulmonary  tuber- 
culosis resemble  those  of  a  deficiency  disease.  He  suggests  that 
collecting  of  statistical  data  on  the  relationship  between  the  con- 
sumption and  the  composition  of  the  diet  in  various  countries  would 
be  helpful  in  the  study  of  this  condition. 

Just  how  important  the  diet  is  in  tuberculosis,  is  shown  by  the 
experimental  investigations  of  Weigert  (1527).  He  showed  in  young 
pigs  that  on  a  constant  protein  content,  tuberculosis  develops  more 
frequently  in  the  presence  of  carbohydrates  than  in  that  of  fats. 
Similar  investigations  were  made  by  E.  Thomas  and  Hornemann 
(1528),  who  showed,  in  pigs  infected  with  tuberculosis,  that  the 
animals  manifested  tuberculosis  less  on  protein  and  much  more  on 
carbohydrates,  while  on  fat,  varying  results  were  obtained.  In 


PATHOLOGICAL    CONDITIONS   DUE    TO   LACK   OF   VITAMINES       385 

some  communications  on  tuberculosis,  particular  attention  was  called 
to  the  significance  of  the  vit amines.  Re*non  (1529)  said  that  on 
vitamine-poor  diets,  a  decreased  immunity  towards  tuberculosis  is 
evident,  and  Muthu  (1530)  likewise  believed  in  the  important  role, 
of  the  vit  amines  in  the  development  of  this  disease. 

LEPROSY 

Button  (1531)  recently  stated  that  in  the  development  of  leprosy 
the  diet,  especially  the  vitamines,  plays  an  important  role.  Sir 
Jonathan  Hutchinson  (1532)  believed  that  leprosy  was  associated  in 
some  way  with  fish  consumption.  On  perusing  the  literature,  we 
see  in  an  article  by  Deycke  (1533),  that  there  is  no  basis  for  this 
assumption  except,  perhaps,  the  statement  that  in  the  treatment,  a 
satisfactory  diet  is  of  significance.  Among  other  known  facts,  we 
see  in  a  paper  by  Underbill,  Honeij,  Bogert  and  Aldrich  (1534)  that 
in  leprosy,  certain  chemical  changes  in  the  bones  are  apparent, 
indicating,  perhaps,  a  disturbance  in  the  calcium  and  magnesium 
metabolism.  The  atrophy  of  the  bones  in  leprosy  is  associated 
with  the  large  requirements  of  these  patients  for  calcium.  If  calcium 
is  added,  marked  retention  results.  We  may,  perhaps,  associate  this 
with  the  action  of  vitamine  A.  Vokurka  (1534a)  observed  an 
increase  in  the  number  of  lepers  in  Bosnia  and  Herzogowina  after 
the  war,  and  attributed  it  to  malnutrition.  It  is  obvious  that 
leprosy  occurs  largely  among  the  poorer  people,  but  any  possible 
relationship  with  the  vitamines  has  not  been  demonstrated .  Never- 
theless, it  would  be  worth  while  to  go  further  into  the  question 
of  diet. 

PNEUMONIA 

The  many  investigators  in  the  field  of  experimental  vitamine 
research  have  quite  regularly  reported  the  frequent  occurrence  of 
infections  of  the  respiratory  apparatus  on  diets  poor  in  vitamines. 
Similar  observations  have  been  made  in  man  by  Fleming,  Macaulay 
and  Clark  (I.e.  910)  in  South  Rhodesia.  Great  epidemics  were  noted 
there  on  a  diet,  previously  described,  which  apparently  leads  to  a 
mixed  avitaminosis.  Vaccine  therapy  was  used  without  success,  and 
there  were  686  deaths  out  of  2251  cases  in  1908,  with  considerable 
meningitis  and  tuberculosis.  At  the  same  time,  the  avitaminosis  was 
evident  in  100  out  of  700  laborers.  Here,  when  a  change  was  made 
in  the  diet,  splendid  results  were  obtained.  Not  only  did  the  avitam- 
inosis disappear  but  the  pneumonia  decreased  also. 


CHAPTER  X 

INFLUENCE  OF  NUTEITION  (VITAMINES)  ON  THE  ACTION  OF  SOME 

POISONS  AND  UPON  PATHOLOGICAL  CONDITIONS  OF 

NON-INFECTIOUS  ORIGIN 

The  older  papers  of  Reid  Hunt  have  already  shown  that  changes 
in  the  diet  are  not  without  effect  on  the  toxicity  of  different  drugs. 
Similarly  Reach  (1535)  has  shown  that  the  resistance  towards  the 
cramps-producing  poison,  picrotoxin,  is  greater  in  mice  on  a  meat 
diet  than  on  a  bread  diet  and  he  assumed  that  he  probably  had  to 
deal  with  the  action  of  some  unknown  substances.  Salant  (1536) 
and  Salant  and  Swanson  (1537)  observed  that  young  carrots  had  a 
far  greater  protective  action  against  the  poisonous  effect  of  sodium 
tartrate  in  rabbits,  than  did  old  carrots.  The  same  results  were 
obtained  with  sweet  potatoes  and  carrot  leaves.  On  oats  and  sugar, 
the  toxicity  was  markedly  increased,  and  the  differences  found  were 
not  due  to  increased  diuresis.  Cats  behave  otherwise,  no  difference 
being  apparent;, here  too,  however,  the  effect  of  the  poison  was 
increased  about  40  per  cent  in  starvation. 

ANEMIA,   ETC. 


Here  we  have  the  work  of  Pearce,  Austin  and  Pepper  (1538),  who 
studied  the  influence  of  the  diet  in  anemia,  after  splenectomy.  They 
observed  in  dogs  that  raw  meat  acts  much  better  than  cooked,  and 
they  thought  this  difference  was  due  to  the  vitamine  content.  Mad- 
sen  (1539)  is  of  the  opinion  that  the  vit amines  play  a  part  in  chlorosis 
and  anemia.  Geiling  and  Green  (1539a)  subjected  this  question 
to  experimental  test.  If  rats  were  given  a  diet  poor  in  protein  vita- 
mines  or  salts,  then  the  blood  regeneration  after  a  hemorrhage  was 
markedly  retarded.  Davis,  Hall  and  Whipple  (1540)  produced 
necrosis  of  the  liver  by  chloroform  and  studied  the  effect  of  different 
diets  on  the  regeneration  of  liver  cells.  Muscle  tissue  and  fats  of 
various  sources  were  without  effect  but  feeding  of  liver,  kidney  and 
brain  resulted  favorably. 

Campbell  (1541)  and  van  der  Bogert  (154 la)  thought  that  the 
occurrence  of  adenoids  in  children  is  of  dietetic  origin,  but  we  do  not 


INFLUENCE    OF   NUTRITION   ON   POISONS  387 

know  how  correct  this  assumption  may  be.  The*  significance  of 
vitamines  in  certain  orthopedic  cases  is  based  on  a  better  foundation. 
Schodel  and  Naumwerk  (I.e.  998)  believed  that  scurvy  predisposes 
to  coxa  vara  and  other  conditions,  diagnosed  as  congenital  hip- 
joint  dislocation.  Hess  (I.e.  1022)  saw,  in  one  case  of  scurvy,  a  bone 
affection  similar  to  coxa  vara.  During  the  war,  Hammer  (1542) 
observed  that  some  fractures  healed  with  great  difficulty,  a  dis- 
turbance in  metabolism  being  justifiably  suspected.  Peckam  (1543) 
believed  that  some  deformities  like  scoliosis,  bow-legs,  flat  feet, 
etc.,  are  of  dietetic  origin.  He  thought  that  most  orthopedists  are 
so  occupied  with  the  purely  orthopedic  aspect  of  their  cases  that 
they  pay  absolutely  no  attention  to  the  practical  important  etio- 
logical  factors.  The  presumptions  of  Peckam  are  certainly  partly 
justified  because  of  the  relationship  between  rickets  and  bone 
formation. 

We  shall  now  discuss  two  diseases  which  are  related  more  to  a 
luxury  consumption  than  to  an  insufficient  dietary,  namely,  diabetes 
and  cancer. 

DIABETES 

We  see  already  the  justification  for  the  conception  that  this  disease 
is  related  to  luxury  consumption  in  the  papers  on  the  effect  of  the  war 
diet.  Magnus-Levy  (1544)  observed  that  during  the  war,  there  was 
a  marked  decrease  in  the  number  of  diabetes  cases,  whereas,  before 
the  war,  they  had  been  constantly  increasing.  Gerhardt  (1545)  had 
the  same  experience  in  Wiirzburg.  Two  phases  of  the  diabetes 
question  interest  us  here.  First,  the  possibility  of  an  antidiabetic 
substance  of  a  vitamine  type  in  the  food,  and  second,  the  danger  of 
an  altogether  rigorous  dietary  restriction  in  the  usual  therapy.  The 
first  possibility  was  justified,  to  a  certain  extent,  for  a  specific  action 
on  diabetes  was  ascribed  to  v.  Noorden's  oatmeal  cure.  Even 
Magnus-Levy  (1546)  tried  to  concentrate  this  hypothetical  substance 
by  alcoholic  extraction,  but  without  success.  Boruttau's  (1547) 
experiments  resulted  differently.  He  found  that  pancreas  extracts, 
yeast  and  yeast  extracts  inhibit  the  cleavage  of  glycogen  in  the  iso- 
lated heart,  while  extracts  from  the  peripheral  oat  layers  decreases  the 
sugar  elimination  in  diabetic  dogs  and  in  man.  Similar  results  were 
obtained  by  Rose  (1548)  with  one  of  the  substances  isolated  from  the 
pancreas.  The  composition  of  the  diet,  especially  its  carbohydrates, 


388  THE    VITAMINES 

influences  the  blood  sugar  content  not  only  of  diabetics  but  also  of 
normal  subjects.  This  was  reported  on  by  Jacobson  (1549),  v. 
Moraczewski  (1550)  and  McCay,  Banerjee,  Ghostal,  Dutta  and  Ray 
(1551).  The  latter  described  conditions  in  India  which  develop 
there  on  a  diet  rich  in  carbohydrates,  and  which  gradually  lead  to 
diabetes.  These  conditions  strongly  resemble  experimental  gly- 
cosuria,  which  we  have  already  described  in  pigeon  beriberi. 

Considering  the  question  of  therapy  in  diabetes,  we  see  the  pos- 
sibility of  producing  an  avitaminosis  by  adhering  too  closely  to  a 
strict  diet.  Burger  (I.e.  1452)  and  Schittenhelm  and  Schlecht  (I.e. 
1443)  state  that  in  diabetes,  on  an  oatmeal  diet,  they  observed 
edema  resembling  hunger  edema.  A  similar  observation  was  made 
by  Wilder  and  Beeler  (155 la).  An  editorial  in  the  Journal  of  the 
American  Medical  Association  (1552)  warned  of  the  dangers  of  a 
decrease  in  the  protein  of  the  diet  with  a  simultaneous  increase  of  the 
carbohydrates,  a  regime  which  produces  real  diabetics  from  latent 
cases.  In  addition,  there  is  the  actual  danger  of  an  avitaminosis. 
As  regards  the  starvation  therapy  in  diabetes,  we  have  asked  our- 
selves the.  question  as  to  whether  the  organism,  in  starvation,  suffers 
first  from  a  lack  of  vitamine  or  of  the  usual  dietary  constituents. 
The  author  and  Dubin  (unpublished  data)  undertook  to  test  this 
question  experimentally.  However,  no  difference  was  noticeable  in 
the  time  of  survival  between  the  pigeons  receiving  either  vitamine  A 
or  B,  or  both,  and  those  that  were  starving.  In  addition,  no  beriberi 
was  observed  in  the  starving  pigeons. 

CANCER 

Cancer  is  a  disease  of  excessive  nutrition,  although  McCarrison 
(1553)  believed  that  under  certain  conditions,  a  lack  of  vitamines 
may  lead  to  cancer  of  the  stomach.  He  saw  one  such  case  among 
monkeys.  That  this  disease  is  associated  with  luxury  consumption, 
is  best  recognizable  from  the  statistical  figures  compiled  during  the 
war  and  from  insurance  statistics.  Hoffmann  (1554)  said  that  cancer 
occurs  more  frequently  in  cities  than  in  the  country,  and  more  among 
the  well-to-do  than  among  poor  people,  the  reverse  being  the  case  in 
tuberculosis. 

The  cancer  question  is  of  interest  from  two  points  of  view,  first,  the 
possibility  of  a  chemical  substance  as  an  etiological  factor  and  second, 


INFLUENCE    OF   NUTRITION   ON   POISONS  389 

the  influence  of  the  diet  on  the  growth  of  neoplasms.  As  for  the 
first  factor,  we  can  not  take  it  up  in  such  great  detail  as  we  should 
like  to,  unfortunately,  since  it  does  not  fall  in  the  sphere  of  this  book. 
We  were  one  of  the  first  to  point  out  this  possibility,  but  we  must 
admit  that  aside  from  the  influence  of  the  diet  on  the  growth  of 
experimental  tumors  and  a  possible  chemical  cause  of  certain  neo- 
plasms, such  as  the  chicken  sarcoma  of  Rous  and  analogous  tumors, 
we  have  no  firm  basis  for  our  conception.  If  we  (1555)  choose  this 
line  of  thought  in  preference  to  the  other  known  ones,  it  is  because 
cancer  research,  in  the  hands  of  pathologists,  has  yielded  very  little 
tangible  results.  Some  of  the  known  pathologists,  like  Ewing  (1556) 
and  Leo  Loeb  (1557)  were  inclined  to  a  similar  view  at  one  time. 
Certain  experimental  papers  like  those  of  Calkins  (1558),  who 
reported  on  the  stimulation  of  protozoa  (Didinium  nasutum)  by 
tumor  extracts,  may  be  interpreted  in  the  above  manner;  still,  it  is 
possible  that  it  may  be  associated  with  the  influence  of  a  dietary  or 
vitamine  addition.  The  various  investigations  dealing  with  the 
effect  of  pregnancy  on  the  growth  of  neoplasms  may  also  be  inter- 
preted from  the  viewpoint  of  the  chemical  theory.  •  From  these 
investigations,  we  may  see  that  the  embryo  contains  a  substance 
that  stimulates  the  growth  of  neoplasms.  The  growing  embryo, 
however,  according  to  v.  Graff  (1559)  and  Slye  (1560),  needs  this 
substance  for  its  own  growth,  thereby  inhibiting  considerably  the 
tumor  growth. 

That  the  diet  exerts  some  influence  on  tumor  growth,  was  suspected 
by  Ehrlich  in  his  athreptic  theory.  This  theory  was  tested  experi- 
mentally by  Jansen  (1561)  and  Haaland  (1562)  who  established  the 
inhibiting  influence  of  under-feeding.  Similar  experiments  were 
also  undertaken  by  Cramer  and  Pringle  (1563),  Rous  (1564)  and 
v.  Jaworski  (1565).  The  latter  believed  that  under-feeding  inhibits 
the  growth  of  tumors.  In  his  opinion,  carcinomas  were  an  exception 
since,  under  such  conditions,  they  grew  even  more  rapidly.  The 
dietetic  aspect  of  cancer  research  was  stimulated  by  vitamine  studies. 
Sweet,  Corson-White  and  Saxon  (1566),  in  their  rat  and  mouse  experi- 
ments, used  the  Osborne-Mendel  diet  and  saw  that  the  growth  of 
tumors  on  a  vitamine-poor  diet  was  markedly  arrested.  In  quick 
succession,  there  appeared  papers  on  the  same  subject  by  Hopkins 
(1567),  Centanni  (1568),  Rous  (1569),  Drummond  (1570)  (this  work 
being  very  exhaustive),  Benedict  and  Rahe  (1571)  and  Sugiura  and 


390  THE   VITAMINES 

Benedict  (1572).  Van  Alstyne  and  Beebe  (1573)  believed  that  they 
had  stimulated  tumor  growth  by  the  addition  of  lactose  to  an  artificial 
diet,  but  this  was  evidently  due  to  a  contamination  of  the  lactose 
with  vitamine  B.  In  a  similar  manner,  the  author  (I.e.  244,  245,  and 
246)  showed  in  1913  on  chickens,  that  the  chicken  sarcoma  of  Rous 
grows  much  slower  on  a  vitamine-poor  diet  than  on  one  rich  in  vita- 
mines,  and  these  results  were  confirmed  by  Drummond  (I.e.  247). 
Levin  (1574),  however,  could  not  corroborate  these  findings  and 
Frankel  and  Fiirer  (1575)  expressed  themselves  against  this  phase  of 
cancer  research.  The  few  negative  results,  naturally,  cannot  have 
any  effect  on  our  conclusions  in  comparison  with  the  conformity  of 
the  conclusions  of  a  great  number  of  investigators  in  this  field. 
Undoubtedly,  the  restriction  of  the  vitamines  in  the  diet  markedly 
inhibits  the  growth  of  neoplasms.  This  measure  would  have  been 
of  practical  value,  if  it  had  not  been  evident  that  the  tumor  tissue 
shows  a  greater  affinity  for  the  vitamines  than  does  the  somatic 
tissue.  This  indicates  that  in  order  to  obtain  a  complete  success, 
the  vitamines  would  have  to  be  so  far  curtailed  that  the  organism 
itself  would  die.  That  certain  substances  govern  tumor  growth,  is 
evident  from  the  paper  by  Rondoni  (1576),  who  showed  that  the 
sarcoma  tissue  could  favorably  influence  the  growth  of  experimental 
tumors.  The  same  conclusions  were  obvious  from  the  work  of 
Murphy  (1577),  who  allowed  rat  tumors  to  grow  in  chicken  embryo, 
it  being  impossible  to  do  so  in  grown  chickens.  From  this,  it  is 
evident  that  in  rapidly  growing  embryonic  tissue,  a  substance  is 
present  which  stimulates  the  growth  of  neoplasms.  That  this  view 
is  justified,  is  clear  from  the  work  of  Funk  (1578),  who  showed  that 
when  the  mouse  chondroma  of  Ehrlich  is  implanted  in  the  rat,  the 
tumor  tissue  is  resorbed  in  a  few  days.  However,  if  the  mouse  tumor 
is  fed  to  the  rat  in  large  quantities,  then  not  only  does  the  tumor 
develop  but  it  may  be  transplanted  in  three  generations  of  rats  so 
treated.  Here,  in  our  opinion,  it  cannot  be  denied  that  the  mouse 
chondroma  contains  a  substance  which  acts  specifically  on  the 
growth  of  a  similar  tumor.  At  the  same  time,  the  influence  of  the 
diet  is  shown  very  well.  The  induced  tumor  possessed,  as  the 
illustration  shows,  for  the  most  part,  the  same  histological  structure. 
The  dietetic  restriction  for  the  purpose  of  treating  cancer  has  also 
been  used  in  practice.  Bulkley  (1579)  used,  for  this  purpose,  a  diet 
so  chosen  that  it  contained  almost  no  animal  protein.  Since  this 


INFLUENCE   OF  NUTRITION   ON   POISONS 


391 


diet  could  be  regarded  for  many  reasons  as  pellagra-producing,  we 
inquired  as  to  the  health  of  the  patients  and  the  duration  of  the 
therapy.  Dr.  Bulkley  replied  that  a  number  of  patients  lived 
on  the  diet  without  showing  any  signs  of  pellagra.  Copeman  (1580) 
tried  a  similar  therapy  with  a  diet  that  was  almost  vitamine-free. 
At  the  time  of  his  preliminary  communication,  his  experiments  had 
lasted  from  three  to  four  months,  and  a  detailed  report  was  still 
to  be  made.  His  patients  increased  in  weight.  The  dietetic  treat- 
ment of  cancer  is  at  present  not  in  favor  with  the  medical  profession. 
We  believe  that  it  would  be  of  value  to  cancer  research  if  this 
disease  were  regarded  more  as  a  metabolic  disturbance. 


>•?    (§M.     •>•;>.>, 

i 


£<&  *  'N» 


A  B 

FIG.  73.  A,  EHRLICH'S  MOUSE  CHONDROMA;  B,  THE  SAME  TUMOR  GROWING  IN 
A  RAT  AFTER  TUMOR  FEEDING 

We  shall  now  consider  two  problems  in  which  the  vitamines  are 
of  significance  with  greater  certainty — the  influence  of  the  diet  in 
the  development  of  teeth,  and  the  cause  of  cystic  calculi  and  similar 
formations. 

THE    DEVELOPMENT   OF   TEETH 

It  was  suspected  by  Kunert  (1581)  that  the  diet  exerted  some 
influence  on  the  condition  of  the  teeth.  Durand  (1582)  saw  a  greater 
percentage  of  carious  teeth  in  children  who  had  been  fed  on  condensed 
milk  than  in  those  fed  on  breast-milk.  A  carbohydrate-rich  diet 
in  particular,  was  held  to  be  responsible  for  caries.  Black  and 


392  THE   VITAMINES 

McKay  (1583)  frequently  observed  a  defective  enamel  in  certain 
districts  in  the  Rocky  Mountains,  where  the  diet  was  inadequate. 
Castilla  (1584)  often  observed  caries  and  defective  enamel  after 
colitis  and  other  disturbances  of  the  stomach  and  intestinal  tract 
in  children  from  1J  to  4  years  old.  Considerable  progress  has  been 
made  in  this  field  since  the  problem  has  been  undertaken  experi- 
mentally. 

Miller  and  Gies  (1585)  studied  the  question  of  teeth  formation  in 
rats  from  the  standpoint  of  the  calcium  and  magnesium  metabolism. 
M.  Mellanby  (1586)  studied  the  condition  of  the  teeth  in  young 
rachitic  dogs.  On  diets  that  were  poor  in  vitamine  A,  the  following 
changes  were  visible : 

1.  Delayed  falling  out  of  the  milk-teeth. 

2.  Delayed  appearance  of  permanent  teeth. 

3.  Disarrangement  in  the  position  of  the  teeth 

4.  Lack  of  or  defects  in  the  enamel. 

5.  Diminished  calcium  content. 

In  quickly  growing  teeth,  the  defects  were  even  more  pronounced. 
Vitamine  A,  in  the  form  of  cod  liver  oil  or  butter,  exerted  a  very 
favorable  influence.  Zilva  and  Wells  (1587)  observed,  in  guinea 
pigs  and  monkeys  on  a  scurvy-producing  diet,  a  fibroid  degeneration 
of  the  tooth  pulp.  These  changes  occur  very  early,  often  before  all 
other  scorbutic  symptoms.  The  findings  on  scorbutic  guinea  pigs 
were  confirmed  by  Percy  R.  Howe  (1588),  Robb,  Medes,  McClendon, 
Graham  and  Murphy  (1588a)  and  developed  further  experimentally 
by  P.  R.  Howe  (1588b).  The  latter  managed,  by  means  of  a  partial 
lack  of  vitamine  C,  to  keep  the  animals  (guinea  pigs  and  rabbits) 
alive  for  about  one  year,  so  that  the  results  were  more  definite. 
The  teeth  were  carious,  poor  in  calcium  and  bent,  with 
Pyorrhoea  alveolaris.  There  was  also  observed  a  disease  of  the 
joints,  resembling  rheumatism  or  Arthritis  deformans.  Ballantyne 
(1589)  found  that  98  per  cent  of  pregnant  women  show  carious  teeth. 
This  may  well  be  explained  by  the  greater  requirements  for  vita- 
mines,  calcium  salts,  etc.  Sinclair  (1590)  rightly  emphasized  that 
precautions  for  the  good  condition  of  the  teeth  of  the  child  must 
begin  with  the  pregnant  mother,  who  should  receive  the  best  possible 
nourishment.  The  effect  of  the  diet  on  the  teeth  does  not  appear 
to  be  related  to  any  one  specific  vitamine,  but  vitamine  A,  because 


INFLUENCE    OF   NUTRITION   ON   POISONS  393 

of  its  relationship  to  calcium  metabolism  seems  to  play  the  most 
important  part.  There  is  no  doubt  that  the  significance  of  the 
factors  mentioned  will  soon  receive  greater  attention  in  practice. 

CALCULI 

Osborne  and  Mendel  (1591)  found  phosphatic  calculi  in  the 
bladder  in  857  rats  kept  on  a  diet  poor  in  vitamine  A.  Padua  (1592) 
saw  a  definite  relationship  between  beriberi  and  calculi  in  the 
Philippines.  Out  of  58  cases  of  phosphatic  calculi,  11  had  beriberi; 
in  18  cases,  under-nutrition  was  observed,  but  without  any  beriberi 
manifestations;  27  cases  occurred  in  well-nourished  patients.  The 
phosphatic  calculi  were  found  mostly  in  children  and  young  people; 
while  urate  calculi  were  observed  later  in  life  (after  50  years). 
Kirschner  (1593)  and  Clemm  (1594)  observed  that  in  Germany,  on 
diets  poor  in  fat,  gall-stones  are  found  more  commonly.  All  these 
cases  are  associated  with  the  indirect  influence  of  the  vitamines, 
presumably  decreasing  the  resistance  to  infections,  in  which  connec- 
tion vitamine  A  is  of  special  significance.  For  it  is  apparent  that  a 
local  infection  is  the  real  cause  of  this  calculus  formation. 


This  concludes  our  data  on  the  subject.  We  are  well  aware  that 
despite  the  ten  years  of  experimental  and  clinical  research,  most  of 
the  problems  discussed  by  us  have  only  been  scratched  on  the  surface. 
This  is  due  largely  to  the  fact  that  such  slow  progress  has  been  made 
in  the  chemistry  of  the  vitamines.  In  this  connection,  the  frequent 
modifications  of  the  nomenclature  is  not  of  the  slightest  value,  and  we 
must  still  await  the  investigator  who,  through  tedious  and  patient 
work,  will  make  the  decisive  step  forward — the  recognition  of  the 
exact  nature  of  the  vitamines. 

Before  closing,  we  shall  add  a  few  words  touching  upon  the  prac- 
tical aspect  of  the  vitamines.  From  what  has  already  been  said, 
it  is  evident  that  in  a  properly  constituted  diet  there  is  no  danger 
of  an  avitaminosis.  In  England  and  in  the  United  States  recently, 
there  have  appeared  on  the  market  a  number  of  vitamine  prepara- 
tions whose  purpose  was  to  exploit  the  present  popularity  of  the 
vitamines.  There  seems  to  be  no  doubt  that  most  of  these  prepara- 
tions cannot  produce  the  effect  claimed  for  them.  Many  investi- 
gators, among  them  Drummond  (1595),  have  expressed  themselves 


394  THE   VITAMINES 

as  being  opposed  to  such  preparations,  maintaining  that  the  natural 
foodstuffs  may  be  used  in  the  therapy  and  the  prevention  of  avitam- 
inoses.  In  the  light  of  our  present  knowledge,  this  appears  indeed 
to  be  the  case.  Still,  we  do  not  know  how  this  matter  will  develop 
in  the  future,  and  the  question  can  be  studied  only  then  when  the 
vitamines  will  be  obtained  in  the  pure  or  highly  concentrated  state. 
It  is  possible  that  such  pure  products  may  manifest  properties  at 
present  neither  known  nor  even  suspected.  For  example,  on  com- 
paring the  influence  on  rickets  of  vitamine  A  of  butter  and  cod  liver 
oil,  it  was  found  that  only  the  latter  can  prevent  the  occurrence  of 
the  disease;  one  might  therefore  be  tempted  to  regard  the  two  sub- 
stances as  individual  chemical  entities.  As  a  matter  of  fact,  if 
it  is  true  that  cod  liver  oil  is  about  250  times  richer  in  vitamine  A 
than  is  butter,  the  above  difference  may  perhaps  be  only 
one  of  concentration.  Although  most  commercial  products  cannot 
withstand  sharp  scrutiny,  it  would  be  desirable  not  to  decry  the 
present  concentration  and  purification  experiments,  since  they  may 
perhaps  lead  to  interesting  and  important  results. 


GENERAL  LITERATURE 

In  order  to  give  the  reader  a  broader  survey  of  the  field,  there  is 
appended  a  chronological  list  of  some  of  the  work  done  on  vitamines 
during  the  last  six  years,  insofar  as  the  records  were  available  and 
insofar  'as  they  will  not  be  referred  to  again  in  the  text.  These 
references  to  the  existing  literature  will  enable  the  interested  reader 
to  make  himself  familiar  with  the  views  of  other  workers.  Reports 
appearing  from  time  to  time  in  the  daily  newspapers  have  not  been 
considered  at  all. 

1914 

FUNK,  CASIMIE:  Die  Vitaminlehre,  ihre  wissenschaftliche  und  praktische 
Bedeutung.  Naturwiss.  2,  121. 

HOPKINS,  F.  G. :  Progress  in  physiological  chemistry.  Ann.  Rep.  Chem. 
Soc.  11,  188. 

HUSSY,  P. :  Zur  klinischen  Bedeutung  der  Vitamine.     M.  med.  W.  61,  No.  18. 

MELOCCHI:  Nuovo  orizzonti  sulF  alimentazione  dell'  uomo.    Napoli. 

MENDEL,  LAFAYETTE  B. :  Newer  points  of  view  regarding  the  part  played  by 
different  food  substances  in  nutrition.  J.  A.  M.  A.  63,  819. 

SCHAEFFER,  G. :  Le  be"ribe*ri  et  les  vitamines  de  Funk.  Unproblemede physio- 
logic generale.  Biologica  15  Avr.,  108. 

STERNBERG,  W. :  Diat  und  diatetische  Behandlung  von  Standpunkte  der 
Vitaminlehre  und  von  Standpunkte  der  diatetischen  Kiiche.  Arch. 
Verdauungsk.  20,  200. 

1915 

BORUTTAU,  H. :  Vitamine  und  akzessorische  Nahrungsbestandteile.     D.  med- 

W.  41,  1208. 
OSBORNE,  T.B.,  AND  MENDEL:  Report  on  Nutrition.   Washington.    Year  Book 

Cam.  Inst.  14,  378.    Also  next  year  15,  365. 
STILES,  PERCY  G. :    The  vitamines.  The  recognition  of  essential  constituents 

of  the  diet  hitherto  unclassified ;  deficiency  diseases.    Am.  J.  Pharm. 

86,  237. 
SCHAUMANN:  Neue    Ergebnisse    der    Beriberiforschung.     Arch.     f.     Schiffs. 

Tropenhyg.,  19,  425,  1915;  Therap.  monatsh.,  29,  152,  1915. 
WATSON-WEMYSS,  H.  L. :  Recent  work  on  vitamines.     Edinb.  Med.   J.,    14, 

186,  1914;  J.  Comp.  Path.  Ther.,  28,  53. 

1916 

ALSBERG,  CARL  L. :  Biochemical  analysis  of  nutrition.     J.  Wash.  Acad.  Sci., 

6,  269. 
FISCHER,  Louis :    Notes  on  the  role  of  vitamines  in  deficiency  diseases.    Amer. 

Med.,  11,  774. 

395 


396  THE  VITAMINES 

FUNK,  CASIMIR:  The  vitamines,  their  chemical  nature,  their  importance  in 

metabolism  and  their  function  in  the  animal  organism.    Amer. 

Med.,  11,  751. 
JACOBS,  MERKEL  H. :  Some  biological  and  practical  aspects  of  growth.  Univers. 

Penn.  Bull.,  17,  Ser.  No.  1,  Part  2,  131. 
McCoLLUM,  E.  V. :  Malnutrition  through  errors  in  the  combination  of  foods. 

Amer.  Food  J.,  11,  404. 

Mosc,  G. :  Deficiency  diseases.    L'ospedale  maggiore  4,  II,  735. 
VEDDER,  E.  B. :  Relation  of  diet  to  beriberi,  and  the  present  status  of  our 

knowledge  of  the  vitamines.     J.  A.  M.  A.,  67,  1494,  1916. 
VOEGTLIN,  C. :  Importance  of  vitamines  in  relation  to  nutrition  in  health  and 

disease.     J.  Wash.  Acad.  Sci.,  6,  575;  The  chemical  nature  and 

physiological  significance  of  so-called  vitamines.    Science  Monthly, 

2,  289. 

1917 

ALB,  B. :  Les  vitamines  et  les  maladies  par  carence.  Revue.  Sci.,  16  et  23  Juin. 
CHRISTENSEN,  F.  W. :  The  so-called  "vitamines"  or  "food  accessories"  and 

their  importance  in  the  nutrition  of  animals.     Special  Bull.  N. 

Dakota  Agr.  Exp.  Sta.,  4,  331. 
CORNALBA,  G. :  Vitamines  and  new  ideas  regarding  human  nutrition.     Boll. 

chim.  farm.,  56,  577. 
McCoLLUM,  E.  V. :  Supplementary  dietary  relationship  among  our  natural 

foodstuffs.     J.  A.  M.  A.,  68,  1379.     Some  essentials  to  a  safe  diet. 

Ann.  Amer.  Acad.  Polit.  and  Soc.  Sci.,  74,  95. 

POL,  D.  J.  H. :  Deficiency  diseases.    Nederl.  Tijdsr.  v.  Geneesk.,  2,  1212. 
STEPP,  WILHELM  :  Einseitige  Ernahrung  und  ihre  Bedeutung  f  iir  die  Pathologic. 

Erg.  d.  inn.  Med.  u.  Kinderh.,  15,  257. 

1918 

ARON,  H. :  Akzessorische  Nahrungsbestandteile.     Berl.  klin.  W.,  55,  546. 
EIJKMAN,   C. :  The  importance  of  vitamines  in  human    nutrition.     Chem. 

Weekbl.,  55,  765. 

KLEISSEL,  R. :  Vitamine.     Wien.  klin.  W.,  68,  601;  Wien.  med.  W.,  p.  547. 
McCoLLUM,  E.  V. :  What  to  teach  the  public  regarding  food  values.     J.  Home 

Econ.,  10,  195;  Influence  of  heat  on  growth-promoting  properties 

of  food.     Amer.  J.  Publ.  Health.,  8,  191. 
RAMSDEN,  W. :  Vitamines.  J.  Soc.  Chem.  Ind.,  37,  53. 
SCHAEFFER,  G.  i  Les  recents  travaux  sur  les  besoins  qualitatifs  d'azote  chez 

les  mammife'res  et  les  vitamines.     Bull.  soc.  sci.  d'hyg.  aliment. 

6,  No.  5  and  6. 

STEENBOCK,  H. :  Vitamines  and  nutrition.     Science  Monthly,  7,  179. 
WEILL,  E.,  AND  MOURIQUAND,  G. :  A  propos  des  vitamines  et  des  maladies  par 

carence.     Rev.  Sci.,  5-12  Juin. 

1919 

Azzi,  A.:  Vitamines  and  oxymones.     Riv.  Med.,  35,  878. 
BUHRER,  C.:  Vitamine.     Schweiz.  Apoth.  Ztg.,  57,  377. 


GENERAL   LITERATURE  397 

COMBY,  J. :  Deficiency  diseases.     Arch,  de  me'd.  d.  enfants.,  22,  659. 

EDDY,  WALTER  H. :  Bibliographic  Review.     The  Vitamines.     Abstr.  Bact., 

3,  313. 
FORBES,  E.  B. :  Vitamines  in  human  nutrition.     Monthl.  Bull.  Ohio  Agr. 

Exp.  Sta.,  4,  299. 
G6MEZ,  A. :  Hormones,  vitamines  and  symbiotes.     Repert.  de  med.  y  cirurgia, 

Bogota,  10,  463. 

GOODBY,  SIR  K:  Vitamines.     Practit.,  103,  6. 

GUIDI,  G. :  Recent  literature  on  vitamines.     Riv.  clin.  pediatr,     17,  243. 
HALLIBURTON,  W.  D. :  Vitamines.     B.  Med.  J.,  May  10,  583. 
HOPKINS,  F.  G. :  Practical  importance  of  vitamines.     Ibid.  Apr.  26th,  507. 
JACOB  Y,   MARTIN:  Die  Bedeutung  der  Qualitat  der  Ernahrung.    Ther.  d.. 

Gegenw.,  60,  401,  1919. 
McCoLLUM,  E.  V. :  Food  control  from  the  standpoint  of  nutrition.    Amer. 

Food  J.,  14,  27,  30.     The  newer  knowledge  of  nutrition.     New  York, 

Macmillan. 
PUGLIESE,  A. :  The  vitamine  problem.    Rend.  Reale.  Inst.  Lomb.  sci.  lett. 

52,  723. 

RISQUEZ:  Vitamines  and  symbiotes.     Gaceta  me'd.  de  Caracas.,  26,  29,  82. 
SCHAEFFER,  G. :  Facteurs  accessoires  de  la  croissance  et  de  Pe*quilibre.     Vita- 
mines,  auximones.  Bull.  Inst.  Past.,  No.  1-2,  17,  1,  41. 
WEILL,  E.,  AND  MOURIQUAND.  G. :  L'alimentation  et  les  maladies  par  carence. 

J.  B.  Bailliere,  Paris. 
WOLFF,  L.  K. :  A  study  of  the  inorganic  constituents  of  the  animal  body  and 

their  significance  as  a  part  of  the  diet.     Chem.  Weekbl.,  16,  1083. 

1990 

AULDE,  J. :  Applied  calcium  therapy.     Study  of  deficiency  diseases.    Med. 

Rec.,  97,  257. 
BLUNT,  K.,  AND  CHI  CHE  WANG:  The  present  status  of  vitamines.     J.  Home 

Econ.,  12,  No.  1,  Jan. 
VAN  DRIEL,  B.:  Present  status  of  vitamine  question.     Nederl.  Tijdsr.  v. 

Geneesk  k,  1350,  Apr.  17. 
FORBES,  E.  B. :  Mineral  nutrients  and  vitamines  in  diet.    Ohio  State  Med. 

J.,  16,  419. 
HAAS,  P.:  Recent  advances  in  science — organic  chemistry.    Sci.  Progr.,  14, 

378. 

HOJER,  A. :  Recent  research  on  the  vitamines.     Hygiea,  82,  449. 
LECOQ,  R. :  Les  nouvelles  theories  de  la  nutrition.    Bull.  Sci.  pharm.,  27,  139. 
LEMAIRE,  H. :  Nourisson,  8,  289. 
LINOSSIER,  G. :  Role  of  different  food  elements  in  nutrition.     Paris  Me'd., 

10, 349. 

LINTON,  R.  J. :  Vitamines.  Science  and  Industry  ( Australia),  2,  438. 
McCLENDON,  J.  F. :  Nutrition  and  public  health,  with   special  reference  to 

vitamines.     Am.  J.  Med.  Sci.,  159,  477. 
McCoLLUM,  E.   V.:  Nutrition  and  physical  efficiency.     J.    Franklin  Inst., 

189,  421. 


398  THE    VITAMINES 

MOLINARI,  G. :  Deficiency  phenomena.     Riv.  med.,  36,  182. 

POL,  D.  J,  HULSHOFF:  One-sided  diet  and  the  vitamines.     Ned.  Tijdsr.  v. 

Gen.,  1,  1625. 
PORTIER,  P.,  Vitamines  and  their  mode  of  action.     Bull.  Soc.   sci.  hyg.  8, 

521,  and  603. 

RAMOINO  :  Policlin.    28,  819. 

SEIDELL,  A.:  Chemistry  of  the  vitamines.     J.  Ind.  Eng.  Chem.,  13,  72,  1921. 
TSCHIRCH,  A. :  What  are  vitamines?    Schweiz.  med.  W.,  50,  21. 
CHICK,  HARRIETTS:    Role  of  vitamines  in  nutrition.    Wien  med.  W.,  70,  411, 

1920. 

NEPPI,  BICE:  Vitamines.     Giorn.  chim.  ind.  appl.,  2,  573,  1920. 
HALLIBURTON,  W.  D.:  Vitamines.     Scientia,  27,  55,  1920. 
RAMSDEN,  W.:  Vitamines.    Dent.  Rec.,  40,  281,  1920. 

1921 

ASHER,  L! :    Review  of  the  vitamine  theory  to  date.    Deutsch.  med.  W.  47, 510. 
BIDAULT:    Vitamines  and  the  preservation  of  food.    Revue  sci.  59,  13. 
BOTAZZI,  FIL.:    The  problem  of  vitamines  and  avitaminoses.    Bull.  soc.  hyg. 

9,1. 
DALYELL,  E. :    Accessory  food  factors  and  their  influence  on  nutrition.    Med. 

J.  of  Australia.  1,61. 

EDDY,  W.  H. :    The  Vitamine  Manual.    Williams  and  Wilkins,  Baltimore. 
EMMETT,  A.  D.:    Physiological  value  of  the  vitamines.    J.  Amer.  Pharm. 

Assocn.  10,  176. 
FITCH,  W.  E.:    The  necessity  for  vitamines  in  the  dietary  and  the  role  they 

enact  in  constructive  metabolism.    Amer.  Med.  27,  368. 
FUNK,  CASIMIR:    Vitamines  and  the  avitaminoses.     Proc.  New  York  Path. 

Soc.  20,  119. 
HAAS,   P.:    Recent  advances  in  science.    Organic  chemistry.     Sci.  Progr. 

15,  377. 
HALLIBURTON,  W.  D.:    Importance  of  the  infinitely  little.     Brit.  Med.  J.  I., 

449;  Lane.  I,  627. 
HARDEN,  A.:    Vitamine.    Chem.  and  Drugs.  94,  202;  Vitamines  and  the  food 

supply.    J.  Soc.  Chem.  Ind.  40,  79. 
HARROW,  B.:    Vitamines.    Essential  food  factors.    Dutton  and  Co.,  New 

York. 
HOPKINS,  "F.  G.:    Recent  advances  in  science  in  their  relation  to  practical 

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MCCARRISON,  R.:    Studies  in  deficiency  diseases.    Oxford  Univer.  Press. 
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Chemical  Monographs.:  Amer.  Chem.  Soc. 
SPERONI,  C.  E. :    Vitamines,  Semana  med.  28,  679. 


LITERATURE  TO  THE  TEXT1 

1.  THOMAS,  K.:  Pfl.  Arch.  291,  1909. 

2.  SCHAUMANN,  H.  i  Die  Aetiologie  der  Beriberi  unter  Beriicksichtigung 

des  gesamten  Stoffwechsels.  Arch.  f.  Schiffs-  u.  Tropenhyg.  14, 
Beih.  8,  1910. 

3.  McCoLLUM,  E.  V.,  HALPIN,  I.  G.  AND  DRESCHER,  A.  H. :  Synthesis  of 

lecithin  in  a  hen  and  the  character  of  the  lecithin  formed.  J.  Biol. 
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4.  FINGERLING:  Die  Bildung  von  organischen  Phosphorverbindungen  aus 

anorganischen Phosphaten.  Biochem.Z.  38,448, 1912; Einfluss  organ- 
ischer  und  anorganischer  Phosphorverbindungen  auf  die  Milchsekre- 
tion.  Ibid.  39,  239,  1921. 

5.  ABDERHALDEN,   E. :  Synthese  der  Zellbausteine  in  Pflanze  und  Tier. 

Berlin  1912.     Springer. 

6.  STEPP,  W. :  Experimentelle  Untersuchungen  liber  die  Bedeutung  der 

Lipoide  fur  die  Ernahrung.     Biochem.  Z.  22,  452,  1909. 

7.  STEPP,  W.:  Z.  f.  Biol.  57,  135,  1911. 

8.  FORSTER,  J. :  Ueber  die  Bedeutung  der  Aschenbestandteile  in  der  Nahrung. 

Z.  f.  Biol.  9,  297,  369,  1873. 

9.  BUNGE,  G. :  Lehrbuch  der  physiologischen  und  pathologischen  Chemie. 

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Ernahrung  des  Tieres.  H.  5,  31,  1881. 

11.  SOCIN,  C.  A. :  In  welcher  Form  wird  das  Eisen  resorbiert?  H.  15,  93,  1891. 

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in  Organismus  des  Sauglings.  H.  16,  173,  1892. 

13.  HAUSERMANN:  Die  Assimilation  des  Eisens.    H.  23,  555,  1897. 

14.  HALL,  WINFRED  S. :  Einige  Bemerkungen  iiber  die  Herstellung  eines 

kiinstlichen  Futters.  Arch.  f.  (Anat)  Physiol.  p.  49,  142,  1896; 
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15.  COPPOLA,  F. :  Sul  valore  fisiologico  e  terapeutico  del  fefro  inorganico. 

Rendic.  R.  Acad.  d.  Line.  6,  I,  362,  1890. 

16.  PASQUALIS,  G. :  Studi  sulla  nutritione  animale.    Atti.  d.  R.  Inst.  Veneto 

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H.  60,  105,  1909. 

18.  FALTA,  W.  AND  NOEGGERATH,  C.  F. :  Fiitterungsversuche  mit  kiinstlicher 

Nahrung.  Hofm.  B.  7,  313,  1905. 

19.  JACOB,   LUDWIG:  Fiitterungsversuche  mit  einer  aus  einfachen    Nahr- 

ungsstoffen  zusammengesetzter  Nahrung  an  Tauben  und  Ratten. 
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20.  MCCOLLUM,  E.  V. :  Am.  J.  Physiol.  25,  120,  1909. 


*For  Index  to  Sectional  Divisions  of  the  Literature,  see  page  476. 

399 


400  THE    VITAMINES 

21.  WATSON,  CHALMERS  D. :  Influence  of  excessive  meat  diet  on  growth,  on 

nutrition  and  on  the  osseous  system.     Effect  of  a  meat  diet  on  the 
animals  and  their  progeny.     Lane.  II.  145,  1585,  1778,  1906. 

22.  WILCOCK,  E.  G.,  AND  HOPKINS,  F.  G. :  The  importance  of  individual 

aminoacids  in  metabolism.     J.  Physiol.  35,  88,  1906-07. 

23.  HOPKINS,  F.  G. :  Analyst  31,  395,  1906. 

24.  ROHMANN,  F. :  Ueber  ktinstliche  Ernahrung  und  Vitamine.     Borntrager 

Berlin,  1916. 

25.  ABDERHALDEN  AND  LAMPE:  Gibt  es  lebenswichtige,  bisher  unbekannte 

Nahrungstoffe?    Z.  ges.  exp.  Med.  1,  296,  1913. 

26.  ABDERHALDEN,  EMIL:  Lehrbuch    der  physiologichen    Chemie.     Urban 

und  Schwarzenberg. 

27.  ABDERHALDEN,  EMIL  AND  FUNK,  CASIMIR:  Zur  Frage  nach  der  Neubil- 

dung  von  Aminosauren  im  tierischen  Organismus.     H.  60,  418,  1909. 

28.  CHITTENDEN,  R.  H.  AND  UNDERBILL,  F.  P. :  Production   in  dogs  of  a 

pathological  condition  which  closely  resembles  human  pellagra. 
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29.  ABDERHALDEN,  EMIL:  Weitere  Studien  liber  Stickstoffwechsel.    H.  96, 

1,  1915. 

30.  GRAFE,  E.,  SCHLAPFER  V.,  TURBAN,  R.  AND  WINTZ,  H.:  H.  77,  1,  1912; 

78,  485,  1912;  83,  25,  1913;  84,  69,  1913;  86,  283,  1913. 

31.  ROHMANN,  F.:  Allg.  med.  Central  Ztg.  1,  1903;  9,  1908. 

32.  KRAMER:  Medicina  castrensis.     1720. 

33.  BACHSTROM:  Observationes  circa  scorbutum.  eiusque  indolem,  causas, 

signa  et  curam.     1734. 

34.  LIND:  A  treatise  on  scurvy.     London  2nd  Edit.,  1757. 

35.  BARLOW,  T. :  Infantile  scurvy.    Brit.  Med.  J.  Nov.  10th,  1894. 

36.  HOLST,   AXEL    AND    FROLICH,  T. :    Experimental    studies    relating   to 

ship-beriberi  and  scurvy.     J.  of  Hyg.  7,  634,  1907. 

37.  MILLER,  R. :  The  medical  diseases  of  children.     New  York,  1911. 

38.  SCHABAD,  J.  A.  AND  SoROCHOWETZ,  R.  F. :  Lipanin,   als    Ersatzmittel 

des  Lebertrans  bei  Rachitis.    Sein  Einfluss  auf  den  Stoffwechsel. 
Monats.  f.  Kinderh.  9,  Orig.  659,  1911. 

39.  SCHABAD   AND   SOROCHOWETZ:  Zur   Frage   von   Wesen   der    gunstigen 

Wirkung  des  Lebertrans  bei  Rachitis.     1st  das  wirksame   Agens 
des  Lebertrans  ein  Ferment?    Ibid.  11,  Orig.  3,  4,  1912. 

40.  WERNICH:  Geographisch-medizinische  Studien.    Berlin,  1878. 

41.  VAN  LEENT:  Gen.  Tijdsr.  voor  Ned.  Indis.,  1880. 

42.  TAKAKI:  Sei-i-kwai  1885,  1886,  1887. 

43.  VORDERMANN:  Gen.  Tijdsr.  voor  Ned.  Indie,  48,  1898. 

44.  BRADDON,  L. :  The  cause  and  prevention  of  beriberi.     London,  1907. 

45.  FLETCHER,  W.:  Rice  and  beri-beri.     Lane.  29th  June,  1907;  J.  Trop. 

Med.  and  Hyg.  12,  No.  9,  p.  127. 

46.  ELLIS,  W.  GILMORE  :  Uncured  rice  as  a  cause  of  beri-beri.    Brit.  Med. 

J.  p.  935,  1909. 

47.  FRASER,  HENRY  AND  STANTON,  H.  T. :  An  inquiry  concerning  the  etiology 

of  beri-beri.     Lane.  76,  451,  1909. 


LITERATURE    TO   THE    TEXT  401 

48.  EIJKMAN,    C. :  Ein    Versuch    zur    Bekampfung    der    Beriberi.     Virch. 

Arch.  149,  187,  1897. 

49.  FRASER,   H.,    AND   STANTON:  An   inquiry   concerning  the   etiology  of 

beriberi.  Studies  from  the  Inst.  for  Med.  Res.,  Feder.  Malay 
States,  No.  10,  1909;  The  etiology  of  beriberi.  Ibid.  No.  12,  1911. 

50.  EIJKMAN,  C. :  Polyneuritis  bij  hoenders,  nieuwe  tot  de  aetiologie  der 

ziekte.  Gen.  Tijdsr.  voor  Ned.  Indie  1896;  Eine  beriberiahnliche 
Krankheit  der  Huhner.  Virch.  Arch.  148,  523,  1897. 

51.  EIJKMAN,  C. :  Ernahrungspolyneuritis.     Arch.  d.  Hyg.,  58,  150,  1906. 

52.  GRIJNS:  Gen.  Tijdsr.  voor  Ned.  Indie  41,  1901;  49,  1909. 

53.  EIJKMAN,  C. :  Virch.  Arch.  222,  301,  1916. 

54.  BELAUD  AT:  Origine  alimentaire  et  traitement  du  be"ribe"ri.    Bull,  de  la 

soc.  de  path.  exot.  p.  13,  1901. 

55.  FRASER  AND  STANTON:  Lane.  p.  4515,  1900. 

56.  SCHUFFNER   AND   KuENEN:  Ueber  den   Einfluss   der  Behandlung  des 

Reises  auf  die  Beriberi  und  die  daraus  entstehenden  Fehlerquellen 
bei  der  Beobachtung  Arch.  f.  Schiffs.  u.  Tropenhyg.  Beih.  7, 
1912. 

57.  SCHATJMANN,  H. :  Beriberi  und  Nukleinphosphorsaure  in   der  Nahrung. 

Arch.  f.  Schiffs.  u.  Tropenhyg.,  12,  Beih.  5,  1908. 

58.  SIMPSON  AND  EDIE  :  On  the  relation  of  the  organic  phosphorus  content 

of  various  diets  to  diseases  of  nutrition,  particularly  beriberi. 
Ann.  Trop.  Med.  a.  Parasit.  5,  313,  1911. 

59.  POL,    HULSHOFF:    Beriberiforschungen     in    den     niederlandisch-ostin- 

dischen  Kolonien,  besonders  in  Bezug  auf  Prophylaxis  und  Heilung 
Arch.  f.  Schiffs-  u.  Tropenhyg.  14.  Beih.  3,  1910. 

60.  THOMPSON  AND  SIMPSON:  Treatment  of  beriberi.    Ann.  Trop.  Med.  a. 

Parasit.  6,  53,  1921. 

61.  EIJKMAN,  C. :  Polyneuritis  gallmarum  and  Beriberi.    Arch.  f.  Schiffs-  u. 

Tropenhyg.  15,  698,  1911. 

62.  FUNK,  CASIMIR:  The  etiology  of  the  deficiency  diseases.     J.  State  Med. 

June,  1912. 

63.  TERUUCHI  :  Ueber  die  Aetiologie  der  Beriberi  unter  besonderer  Beriick- 

sichtigung  der  Schaumann'schen  Phosphormangel  theorie.  Saiking- 
akuzashi.  Tokyo  No.  79,  1910. 

64.  CHAMBERLAIN  AND  VEDDER:  A  contribution  to  the  etiology  of  beriberi. 

Philipp  J.  of  Sci.  6  B,  251,  1911;  6,  395,  1911;  with  Williams,  R.  R., 
Ibid.  7,  39,  1912. 

65.  SHIGA  AND  KUSAMA:  Arch.  f.  Schiffs-  u.  Tropenhyg.  Beih.  3,  May,  1911; 

Shiga:  Experimentelle  Studien  iiber  Kakke.  C.  f.  Bakt.  I  Abt. 
Beih.  6,  Tagung  d.  Vereins  f.  Mikrobiologie  p.  158. 

66.  TSUZUKI  :  Antiberiberintherapie  der  Beriberikrankheit  1912. 

67.  ARON  AND  HOCSON:  Phosphorus  starvation  with  special  reference  to 

beriberi.     II.  Philip.  J.  of  Sci.  5,  February,  1910. 

68.  COOPER  AND  FUNK,  CASIMIR:  Experiments  on  the  causation  of  beriberi. 

Lane.  p.  1266,  1911. 


402  THE    VITAMINES 

69.  McCoLLUM,  E.  V.  AND  PITZ,  W. :   The  "vitamine"  hypothesis  and  defi- 

ciency diseases.     A  study  of  experimental  scurvy.     J.  Biol.  Chem., 
31,  229,  1917. 

70.  HOPKINS,  F.  G. :  Feeding  experiments  illustrating  the  importance  of 

accessory  factors  in  normal  dietaries.     J.  Physiol.  44,  425,  1912. 

71.  OSBORNE,  T.  B.  AND  MENDEL,  L.  B. :  Feeding  experiments  with  isolated 

food-substances.     Publ.  Cam.  Inst.,  Wash.,  No.  156,  Part  I  and  II. 

72.  The  same.     Feeding  experiments  with  fat-free  food  mixtures.     J.  Biol. 

Chem.  12,  81,  1912;  H.  80,  307,  1912. 

73.  The  same.    Maintenance  experiments  with  isolated  proteins.     J.  Biol. 

Chem.  13,  233,  1912. 

74.  HOPKINS  AND  NEVILLE:  A  note  concerning  the  influence  of  diets  upon 

growth.     Biochem.   J.  7,  97,  1913. 

75.  McCoLLUM  AND  DAVIS,  M. :  The  necessity  of  certain  lipins  in  the  diet 

during  growth.     J.  Biol.  Chem.  15,  67,  1913. 

76.  OSBORNE  AND  MENDEL:  The  relation  of  growth  to  the  chemical  constit- 

uents of  the  diet.     J.  Biol.  Chem.  15,  311,  1913. 

77.  The  same.     The  influence  of  butter-fat  on  growth.     Ibid.  16,  423,  1913. 

78.  McCoLLUM  AND  KENNEDY.     Ibid.  24,  491,  1916. 

79.  DRUMMOND,  J.  C. :  Note  on  the  role  of  the  antiscorbutic  factor  in  nutri- 

tion.   Biochem.  J.  13,  77,  1919. 

80.  FUNK,  CASIMIR:  What  is  a  vitamine?    J.  A.  M.  A.  66,  1650,  1916. 

80a.  DRUMMOND,  J.  C. :  The  nomenclature  of  the  so-called  accessory  food 
factors  (vitamines).     Biochem.  J.  14,  660,  1920. 

81.  FUNK,  CASIMIR  AND  MACALLUM,  A.  B.,   JR.:  Die   chemischen   Deter- 

minanten  des  Wachtsums.     Ztschr.  f.  Physiol.  Chem.  92,  13,  1914. 

82.  McCoLLUM  AND  DAVIS  i  Observations  on  the  isolation  of  the  substance 

in  butter-fat  which  exerts  a  stimulating  influence   on   growth.     J. 
Biol.  Chem.  19,  245,  1914. 

83.  OSBORNE  AND  WAKEMAN:  Does  butter-fat  contain  nitrogen  and  phos- 

phorus?   Ibid.  21,  91,  1915. 

84.  WRAMPELMEYER,   E. :  Der  Lecithingehalt  der  Butter.     Landw.   Vers. 

Sta.  42,  437,  1892. 

85.  SUPPLEE,  G.  C. :  The  lecithin  content  of  butter  and  its  possible  relation- 

ship to  the  fishy  flavor.     Corn.  Univers.  Agr.  Exp.  Stat.  29,  101, 
1919. 

86.  FUNK,  CASIMIR  AND  MACALLUM,  A.  B.,  JR.:  Studies   on  growth.     II. 

On  the  probable  nature  of  the  substance  promoting  growth  in 
animals.     J.  Biol.  Chem.  23,  413,  1915. 

87.  DANIELS,  AMY  L.  AND  LOUGHLIN,  ROSEMARY:  Note  on  the  fat-soluble 

growth-promoting  substance    in  lard  and  cotton-seed  oil.     Ibid. 
42,  359,  1920. 

88.  SEIDELL,    ATHERTON:  Vitamines    and   nutritional    diseases;    a    stable 

form  of  vitamine.     U.  S.  Publ.  Health  Rep.,  Repr.,  325,  1916. 

89.  HARDEN,  A.  AND  ZILVA,  S.  S. :   Differential  behavior  of  the  antineuritic 

and  antiscorbutic  factors  towards  adsorbents.     Biochem.  J.  12,  93, 
1918. 


LITERATURE   TO    THE    TEXT  403 

90.  BYFIELD,  A.  H.,  DANIELS,  A.  L.  AND  LOUGHLIN,  R. :  The  antineuritic 

and   growth-stimulating  properties  of   orange  juice.     Am.    J.   Dis. 
Childr.  19,  349,  1920. 

91.  FURST,  VALENTIN:  Weitere  Beitrage  zur  Aetiologie  des  experiment e lien 

Skorbuts  des  Meerschweinchens.  Z.  f.  Hyg.  u.  Infektionsk.  72, 
121,  1912. 

92.  OSBORNE   AND   MENDEL:  Do   fruits   contain  water-soluble    vitamine? 

Soc.  Exp.  Biol.  Med.  17,  46,  1919;  J.  Biol.  Chem.  42,  465,  1920. 

93.  MITCHELL,  H.  H. :  On  the  identity  of  the  water-soluble  growth-pro- 

moting vitamine  and  the  antineuritic  vitamine.  J.  Biol.  Chem. 
40,  399,  1919. 

94.  EMMETT,  A.  D.  AND  LUROS,  G.  O. :  Are  the  antineuritic  and  the  water- 

soluble  vitamines  the  same?  Proc.  Soc.  Biol.  Chem.,  J.  Biol. 
Chem.  41,  VII,  1920;  J.  Biol.  Chem.  43,  265,  1920. 

95.  MELLANBY,  EDWARD:  A  further  demonstration  of  the  part  played  by 

accessory  food  factors  in  the  etiology  of  rickets.  Proc.  Physiol. 
Soc.  Dec.  14th,  1918;  J.  Physiol.  52,  LIII,  1919;  An  experimental 
investigation  on  rickets.  Lane.  March  15,  1919. 

96.  HESS,  ALFRED  F.  AND  UNGER,  L.  J. :  The  role  of  fat-soluble  vitamine 

in  the  dietary  of  infants.  Proc.  Soo.  Exp.  Biol.  Med.  17,  49,  1919; 
The  clinical  role  of  the  fat-soluble  vitamine;  its  relation  to  rickets. 
J.  A.  M.  A.  74,  217,  1920. 

97.  McCoLLUM  AND  DAVIS  :  Essential  factors  in  the  diet  during  growth. 

J.  Biol  Chem.  23,  231,  1915. 

98.  FUNK,  CASIMIR  AND  MACALLUM,  A.  B. :  Studies  on  growth.     III.  The 

comparative  value  of  lard  and  butter  fat.     Ibid.  27,  51,  1916. 

99.  HOGAN,  ALBERT  G. :  The  effect  of  high  temperature  on  the  nutritive 

value  of  foods.     Ibid.  30,  115,  1917. 

100.  HARDEN,  A.  AND  ZILVA,  S.  S. :  Accessory  factors  in  the  nutrition  of  the 

rat.     Bio.  J.  12,  408,  1918. 

101.  FUNK,    CASIMIR:  The   nitrogenous   constituents   of   lime-juice.     Ibid. 

7,  81,  1913. 

102.  OSBORNE,  T.  B.,  WAKEMAN,  A.  J.   AND  FERRY,  E.  A.:  Preparation  of 

protein  free  from  water-soluble  vitamine.  J.  Biol.  Chem.  39,  35, 
1919. 

103.  OSBORNE  AND  MENDEL:  Protein    minima  for  maintenance.     J.   Biol. 

Chem.  22,  241,  1915;  20,  351,  1915;  26,  1,  1916;  37,  223,  1919. 

104.  EDELSTEIN,  F.  AND  LANGSTEIN,  L. :  Das  Eiweissproblem  im  Sauglings 

alter,  experimentelle  Untersuchungen  iiber  die  Wertigkeit  der 
Milcheiweisskorper  fur  das  Wachstum.  Z.  f.  Kinderh.  20,  112, 
1919. 

105.  EMMETT,  A.  D.  AND  LUROS,  G.  O. :  Is  lactalbumin  a  complete  protein 

for  growth?  J.  Biol  Chem.  38,  147,  1919;  The  stability  of  lactal- 
bumin toward  heat.  Ibid.  38,  257,  1919. 

106.  MCCOLLUM,  SIMMONDS  AND  PARSONS:  Ibid.  37,  287,  1919. 

107.  EMMETT,  A.  D.  AND  McKiM,  L.  H. :  The  value  of  the  yeast  vitamine  as 

a  supplement  to  a  rice  diet.     Ibid.  409,  1917. 


404  THE    V1TAMINES 

108.  SUGIURA,  K.  AND  BENEDICT,  S.  R.  i  The  nutritive  value  of  the  banana. 

J.  Biol.  Chem.  36,  171,  1918;  40,  449,  1919. 

109.  GOLDBERGER,  JOSEPH  AND  WHEELER,  G.  A. :  The  experimental  produc- 

tion of  pellagra  in  human  subjects  by  means  of  diet.  Hyg.  Labor. 
Wash.  Bull.  120,  Febr.,  p.  7,  1920. 

110.  CLARK,  GUY  W. :  The  properties  and  composition  of  oocytin.     J.  Biol. 

Chem.  35,  253,  1918. 

111.  ROBERTSON,  BRAILSFORD  T. :  On  the  extraction  of  a  substance  from  the 

sperm  of  a  sea-urchin,  which  will  fertilize  eggs.     Ibid.  12,  1,  1912. 

112.  PASTEUR,  Louis:  Note  sur  un  memoir  de  M.  Liebig,  relatif  aux  fer- 

mentations. 

113.  WILDIERS,  E. :  Nouvelle  substance  indispensable  au  deVeloppement  de 

la  levure.    La  Cellule.  18,  313,  1901. 

114.  AMAND,  ABEL:  Le  "Bios"  de  Wildiers  ne  joue  pas  le  role  d'un  contre- 

poison.     Ibid.  20,  225,  1902;  21,  324,  1904. 

115.  DEVLOO,  RENE:  Ibid.  23,  36,  1906. 

116.  PRINGSHEIM,  H.  H. :  Centr.  f.  Bakt.    II  Abt.  16,  111,  1916. 

117.  RUBNER,  MAX:  Die  Ernahrungsphysiologie  der  Hefezelle  bei  der  alko- 

holischen  Gahrung.     Leipzig,  1913. 

118.  VLAHUTA,  E. :  Preparation  of  a  peptone  by  the  decomposition  of  the 

cells  of  beer  yeast  and  the  role  of  this  peptone  in  fermentation. 
Bull.  acad.  Sci.  Roumanie.  3,  123,  1914-15. 

119.  KURONO,  K. :  Significance  of  oryzanin  for  the  nutrition  of  fermentative 

organisms.     J.  Coll.  Agr.  Univers.  Tokyo.    5,  305,  1915. 

120.  BRILL,  H.  C.  AND  THURLOW,  L.  W. :   Alcohol  from  discarded  molasses 

in  the  Philippine  Islands.    Philipp.  J.  Sci.  12,  267,  1917. 

121.  MOUFANG,   E. :  Beschleunigung  der    Gahrung    (durch    getotete    Hefe) 

Allg.  Brau.  Hopfen.  Ztg.  55,  605,  1915. 

122.  SAITO,  K. :  Chemical  conditions  for  the  development  of  the  reproductive 

organs  of  some  yeasts.    J.  Coll.  Sci.  Univ.  Tokyo,  39,  1,  1916. 

123.  BOKORNY,  TH.  :  Bio.  Z.  82,  359,  1917. 

124.  VANSTEENBERGE,  P. :  The  autolysis  of  yeast  and  the  influence  of  prote- 

olysis  on  the  development  of  yeast  and  lactic  bacteria.  Ann. 
Inst.  Past.,  31,  601,  1917. 

125.  LAMPITT,   L.   H. :  Nitrogen  metabolism  in  Saccharomyces    cerevisiae. 

Biochem.  J.  13,  459,  1919. 

126.  ABDERHALDEN,  E.  AND  SCHAUMANN,  H. :  Die  Wirkung  einiger  aus  der 

Hefe  mit  Alkohol  extrahierten  Substanzen  auf  die  Aktivitat  der 
Hefefermente.  Fermentf.  2,  120,  1918. 

127.  ABDERHALDEN  AND  KOHLER,  A. :  Die  Wirkungsweise  einer  in   Alkohol 

loslichen  Substanz,  wahrend  der  alkoholischen  Gahrung  durch 
Hefe  gebildet,  auf  niedrigere  Organismen.  Arch.  ges.  Physiol. 
176,  209,  1919. 

128.  WILLIAMS,  ROGER  J. :  The  vitamine  requirements  of  yeast.    A  simple 

biological  test  for  vitamine.     J.  Biol.  Chem.  38,  465,  1919. 

129.  BACHMANN,  F.  M. :  Vitamine  requirements  of  certain  yeasts.     J.  Biol. 

Chem.  39,  235,  1919. 


LITERATURE    TO    THE    TEXT  405 

129a.  FULMER,  ELLIS  I.,  NELSON,  VICTOR  E.  AND  SHERWOOD,  F.  F. :  The 
nutritional  requirements  of  yeast.  I.  The  role  of  vitamines  in 
the  growth  of  yeast.  II.  The  effect  of  the  composition  of  the 
medium  on  the  growth  of  yeast.  J.  Amer.  Chem.  Soc.  43,  186- 
191,  1921. 

129b.  NELSON,  V.  E.,  FULMER,  E.  I.  AND  CESSNA,  RUTH:  Nutritional  re- 
quirements of  yeast.  III.  Synthesis  of  Vitamine  B  by  yeast. 
J.Biol.Chem.46,77, 1921. 

129c.  HARDEN,  A.  AND  ZILVA,  S.  S.:  The  synthesis  of  vitamine  B  by  yeasts 
(preliminary  note) .  Biochem.  J.  15,  438, 1921 . 

129d.  FUNK,  CASIMIR  AND  DUBIN,  HARRY  E.:  Vitamine  requirements  of 
certain  yeast  and  bacteria.  J.  Biol.  Chem.  48,  437,  1921. 

129e.  EDDY,  WALTER  H.,  HEFT,  HATTIE  L.,  STEVENSON,  HELEN  C.,  AND 
JOHNSON,  RUTH:  Proc.  Soc.  Exp.  Biol.  Med.  18,  138,  1921. 

129f.  MACDONALD,  MARGARET  B.  AND  McCoLLUM,  E.  V. :  The  cultivation 
of  yeast  in  solutions  of  purified  nutrients.  J.  Biol.  Chem.  45,  307, 
1921. 

129g.  IDE,  M.:  The  "Bios"  of  Wildiers  and  the  cultivation  of  yeast.  J. 
Biol.  Chem.  46,  521,  1921. 

129h.  MACDONALD,  M.  B.  AND  McCoLLUM,  E.  V.:  The  "Bios"  of  Wildiers 
and  the  cultivation  of  yeast.  Ibid.  46,  525,  1921. 

129i.  WILLIAMS,  R.  J.:    Vitamines  and  yeast  growth.    Ibid.  46,  113,  1921. 

130.  BERTRAND,  GABRIEL:  Etudes  biologiques  sur  la  bacte"rie  du   sorbose. 

Ann.  de  chim.  et  de  phys.  (8),  3,  121,  1904. 

131.  NOGUCHI,  H. :  A  method  for  the  pure  cultivation  of  pathogenic  Trepo- 

nema  pallidum.    (Spirochaeta   pallida).     J.    Exp.    Med.    14,   99, 
1911. 

132.  VEDDER:  J.  Infect.  Dis.  16,  385,  1915. 

133.  DORYLAND,  C.  J.  T. :  J.  Bact.  1,  135,  1916. 

134.  PIEPER,  E.  J.,  HUMPHREY,  C.  J.AND  ACREE,  S.  F. :  Phytopath.  7,  214, 1917. 

135.  LOCKEMAN,  GEORG:  Welche  Substanzen  sind  zum  Wachstum  des  Tuber- 

kulose  Bacillus  absolut  notig.     Centr.  Bakt.  u.  Parasitenk.     Abt. 
I,  83,  420,  1919. 

136.  BAINB RIDGE.     J.  Hyg.  11,  341,  1911. 

137.  SPERRY  AND  RETTGER:  J.  Biol.  Chem.  20,  445,  1915. 

138.  ROBINSON,  HAROLD  C.  AND  RETTGER,  LEO  F. :  The  growth  of  bacterias 

in  protein-free   enzyme-   and   acid-digestion  products.     J.   Bact. 
3,  209,  1918. 

139.  PACINI,  J.  P.  AND  RUSSELL,  D.  W. :   The  presence  of  growth-producing 

substances  in  cultures    of    typhoid  bacilli.     J.    Biol.    Chem.   34, 
43,  1918. 

140.  MACKENZIE  WALLIS,  H.  L. :  Improvements   in   bacteriological  media. 

I.  A  new  and  efficient  substitute  for  "Nutrose".    Agr.  J.    India 
12,  621,  1917. 

141.  HUNTOON,  F.  M. :  Hormone  medium,  a  simple  medium  employable  as  a 

substitute  for  serum  medium.     J.  Infect.  Dis.  23,  169,  1918. 

142.  AYERS,  S.  H.  AND  RUPP,  P.:  Extracts  of  pure  dry   yeast  for  culture 

medium.     J.  Bact.  5,  89,  1920. 


406  THE    VITAMINES 

143.  LLOYD,    DOROTHY   J. :  Vitamines,    amino    acids,    and    other    chemical 

factors  involved  in  the  growth  of  the  Meningococcus.  J.  Path. 
Bact.  21,  113,  1916. 

144.  The  same.    Brit.  Med.  J.  II,  143,  1916. 

145.  The  same.     Chemical  factors  involved  in  the  growth  of  the  Meningo- 

coccus.    Ibid.  I,  11,  1917. 

146.  GORDON,  M.  H.  AND  HINE,  T.  G.  M. :  Experimental  study  of  the  cultural 

requirements  of  the  Meningococcus.     Ibid.  II,  678,  1916. 

147.  FLACK,   MARTIN:  Note   on   the   constituent   of  pea-flour   augmenting 

growth  of  the  Meningococcus  on  the  trypagar.  medium.  Ibid.  II, 
682,  1916. 

148.  SHEARER,  C. :  The  presence  of  an  accessory  food  factor  in  the  nasal 

secretion  and  its  action  on  the  growth  of  the  Meningococcus  and 
other  pathogenic  bacteria.  Lane.  I,  59,  1917. 

149.  The  same.     The  action  of  spinal  fluid  in  stimulating  the  growth  of  the 

Meningococcus.     Ibid.  II,  714,  1917. 

150.  EBERSON,  FREDERICK:  Yeast  mycelium  for  prolonging  the  viability  of 

Meningococcus.     J.  A.  M.  A.  72,  852,  1919. 

151.  COLE,  SYDNEY  W.  AND  LLOYD,  DOROTHY  J. :  The  preparation  of  solid 

and  liquid  media  for  the  cultivation  of  the  Gonococcus.  J.  Path. 
Bact.  21,  267,  1917. 

152.  MORINI,   L. :  Cultivation  of  Gonococcus  in  gelatin  with  beer  yeast. 

Rif.  med.  Napoli.  36,  2,  1920. 

153.  DOUGLAS,  S.  R.,  FLEMING,  A.  AND  COLEBROOK,  L. :  Studies  in  wound 

infections.  The  growth  of  anaerobic  bacilli  in  fluid  media  under 
apparently  aerobic  conditions.  Lane.  II.  530,  1917. 

153a.  SAZERAC,  ROBERT:    C.  r.  171,  278,  1920. 

153b.  MASUCCI,  PETER:  A  note  on  the  effect  of  amino  acids  on  the  growth 
of  tubercle  bacilli.  J.  Lab.  Clin.  Med.  6,  96,  1920. 

154.  DAVIS,  DAVID  JOHN:  Food  accessory  factors  (vitamines)  in  bacterial 

culture,  with  special  reference  to  hemophilic  bacilli.     J.  Inf.  Dis. 

21,  392,  1917. 

155.  The  same.     II.  Ibid.  23,  248,  1918. 

156.  WOLF,  C.  G.  L.  AND  HARRIS,  J.  E.  G. :  Contribution  to  the  biochemistry 

of  pathogenic  anaerobes.  IV.  The  biochemistry  of  Bacillus 
histolyticus.  J.  Path.  Bact.  22,  1,  1918. 

157.  WOLF,    C.    G.    L.:  V.  The   biochemistry   of   Vibrion    septique.     Ibid. 

22,  115,  1918. 

158.  The  same.     VI.  The  biochemistry  of  Bacillus  sporogenes  and  Bacillus 

Welchii.  VII.  The  biochemistry  of  Bacillus  proteus.  Ibid.  22, 
270,  1919. 

159.  BOYER,  Louis:  Un  nouveau  milieu  de  culture,  particulierement  adapte* 

au  developpement  du  streptococce  pyogene.  C.  r.  soc.  de  biol. 
81,  229,  1918. 

160.  OTABE,  SHORABURO:  Wheat  culture-media.     Lane.  197,  576,  1919. 

161.  DIENERT,  F.  AND  GUILLERD,  A.:  Milieu  de  culture  d'un  extrait  de  la 

levure  autolyse"e  pour  les  cultures  de  B.  coli.     C.  r.  168,  256,  1919. 


LITERATURE    TO    THE    TEXT  407 

162.  BUNKER,  JOHN  W.  M. :  Studies  on  the  diphtheria  bacillus  in  culture. 

J.  Bact.  4,  379,  1919. 

163.  DAVIS,  L.  AND  FERRY,  N.  S. :   Studies  on  diphtheria  toxin  II.     The  role 

of  the  aminoacids  in  the   metabolism  of   B    diphtheriae.      Ibid. 
4,  217,  1919. 

164.  AGULHON,  H.  AND  LEGROTJX,  R. :  Contribution  a  Petude  des  vitamines 

utilisables.  a  la  culture  des  microorganismes.    Application  au  ba- 
cille  de  1' influenza  (B.  de  Pfeiffer)     C.  r.  167,  597,  1918. 

165.  LEGROUX,  REN£  AND  MESNARD,  JOSEPH  :  Les  vitamines  pour  les  cultures 

des  bacteries.     Ibid.  170,  901,  1920. 
165a.  DAVIS,  D.  J.:    Food  accessory  factors  in  bacterial  growth.     Ill,  IV, 

V.    J.  Infect.  Dis.  29,  171,  1921. 
165b.  RIVERS,  T.  M.  AND  POOLE,  A.  K.:    Growth  requirements  of  influenza 

bacilli.    Johns  Hopk.  Hosp.  Bull.  32,  202,  1921. 
165c.  THJOTTA,   TH.:    Studies   in   bacterial  nutrition.    J.   Exp.  Med.  33, 

763,  1921. 
165d.  THJOTTA,  TH.  AND  AVERT,  O.  T.:    Growth  accessory  substances  in 

nutrition  of  bacteria.    Proc.  Soc.  Exp.  Biol.  Med.  18,  197,  1921; 

J.  Exp.  Med.  34,  97,  1921. 

166.  KLIGLER,  I.  J. :  Yeast  autolysate  as  a  culture  medium  for  bacteria.    J. 

Bact.  4,  183,  1919. 

167.  The  same.     Growth  accessory  substances  for  pathogenic    bacteria  in 

animal  tissues.     J.  Exp.  Med.  30,  31,  1919. 

167a.  MUELLER,  J.  HOWARD:  Observations  of  bacterial  metabolism.  Proc. 
Soc.  Exp.  Biol.  Med.  Oct.  20,  1920. 

167b.  MUELLER,  J.  HOWARD,:  Growth  determining  substances  in  bac- 
teriological culture  media.  Soc.  Exp.  Biol.  Med.  Apr.  10,  1921. 

167c.  FREEDMAN,  L.:  Comparison  of  yeast  and  bacteria  growth-promoting 
vitamines.  Amer.  Chem.  Soc.  Sept.  1921. 

167d.  NORRIS,  D. :  Preparation  of  a  simplified  culture  medium  for  field 
work.  Ind.  J.  Med.  Res.  7,  704,  1920. 

168.  LUTZ,  C. :  Ann.  Mycol.  von  Sydow.     7,  91,  1909. 

169.  CURRIE,  JAMES  N. :  The  citric  acid  fermentation  of  Aspergillus  niger. 

J.  Biol.  Chem.     31,  15,  1917. 

170.  WILLAMAN,  J.  J. :  J.  Ind.  Eng.  Chem.     10,  863,  1918;  The  function  of 

vitamines   in   the   metabolism   of   Sclerotinia   cinerea.     J.   Amer. 
Chem.  Soc.  42,  549,  1920. 
170a.  LINOSSIER,  G. :  C.  r.  soc.  biol.    82,  381,  1919. 

171.  CLINTON,  G.  P. :  Conn.  Agr.  Exp.  Sta.  Rep.  1909-10,  p.  753. 

172.  HABERLANDT,    G. :  Zur    Physiologic    der     Zellteilung.    Sitzungsb.    d. 

Preuss.  Akad.  Wiss.  Berlin.  318,  1913. 

173.  BOTTOMLEY,  W.  B. :  The  bacterial  treatment  of  peat.     J.  R.  Soc.  Arts. 

72,  372;  Some  effects  of  humates  on  plant  growth.    Rep.  Br.  Ass. 
Adv.  of  Sciences.  680,  1912. 

174.  The  same.     Some  accessory  factors  in  plant  growth  and  nutrition. 

Proc.  R.  Soc.  88   B,    237,    1914;  The  significance  of  certain  food 
substances  for  plant  growth.     Ann.  Bot.  28,  531,  1914. 


408  THE    VITAMINES 

175.  Anonymous:  Experiments  with  bacterized  peat.     Agr.  News.  13,  263, 

1914. 

176.  BOTTOMLEY,  W.  B.  i  A  bacterial  test  for  plant  food  accessories  (Auxi- 

mones).     Proc.  Roy.  Soc.  89  B,  102,  1915. 

177.  CHITTENDEN,   F.   I. :  Report  on  experiments  with  bacterized  peat  or 

humogen.     J.  Roy.  Hort.  Soc.  41,  305,  1915. 

178.  BOTTOMLEY:  Some    effect    of    organic    growth-promoting    substances 

(auximones),  on  the  growth  of  Lemna  minor  in  mineral  culture 
solution.     Proc.  R.  Soc.  89  B,  481,  1917. 

179.  The  same.     Ann.  Bot.  34,  1920. 

180.  ROSENHEIM,  OTTO:  Accessory  factors  for  plant  growth.     Biochem.  J. 

11,  7,  1917. 

181.  BOTTOMLEY:  Isolation  from  peat  of  certain  nucleic  acid  derivatives. 

Proc.  R.  Soc.  90  B,  39,  1917. 

182.  APPLEMAN,  C.  O.:  Maryland  Agr.  Exp.  Sta.  Bull.    212,  1918;  Science 

N.  S.  48,  319,  1918. 

183.  BOTTOMLEY:  Effect    of    nitrogen-fixing    organisms    and    nucleic    acid 

derivatives  on  plant  growth.     Proc.  R.  Soc.  91  B,  83,  1919. 

184.  JONES,  D.  H.    Growth  of  plants.    Abstr.  Bact.  1,  43,  1917. 

185.  STEAD,  ARTHUR.     Notes  on  humus,  humogen,  and  its  accessory  plant 

food  substances.    S.  Afr.  J.  Sci.  13,  239,  1917. 

186.  MOCKERIDGE,  FLORENCE  A. :  Some  effects  of  organic  growth-promoting 

substances   (auximones)   on  the  soil  organisms  concerned  in  the 
nitrogen  cycle.     Proc.  R.  Soc.  89  B,  508,  1917. 

187.  SCHREINER,  REED  AND  SKINNER:  U.  S.  Dept.  Agr.  Bureau  of  Soils.    Bull. ' 

47,  1907;  Schreiner  and  Skinner:  Ibid.  Bull.  87,  1912;  Schreiner  and 
Shorey:  Ibid.  Bull.  74,  1910;  Shorey  Bull.  88,  1913. 

188.  MACALISTER:   The    action    of    Symphytum    ofncinale    and    allantoin. 

Brit.  Med.  J.  Sept.  21,  1912. 

189.  COPPIN:  The  effect  of  purine  derivatives  and  other  organic  compounds 

on  growth  and  cell-division  in  plants.     Biochem.  J.  6,  416,  1912. 

190.  MOCKERIDGE,   FLORENCE  ANNIE:  The  occurrence  and  nature  of  the 

plant-producing  substances  in  various  mammal  imposts.     Ibid. 
14,  432,  1920. 

191.  GRAFE,  VICTOR:  Sterilisieren  le bender  Pflanzen.     Abderhaldens  Handb. 

d.  biochem.  Arbeitsm.     6,  139,  1912;  8,  178,  1915. 

191a.  PARSONS,  HELEN  T. :  The  antiscorbutic  content  of  certain  body  tissues 
of  the  rat.  The  persistence  of  the  antiscorbutic  substance  of  the 
rat  after  long  intervals  on  a  scorbutic  diet.  J.  Biol.  Chem.  44, 
587,  1920. 

192.  SPRIGGS,  E.  J. :  The  effect  of  a  meat  diet  upon  the  coats  of  rats  with  and 

without  the  supply  of  lime.     J.  Physiol.  36,  xvii,  1907. 

193.  MAIGNON,  F. :  L'influence  des  saisons  sur  la  toxicite  du  blanc  d'oeuf, 

chez  les  rats  blancs.     C.  r.  de  la  ass.  franc,  avanc.  sci.  43,  553;  C.  r. 
166,  1008,  1918;  167,  91,  1918. 

194.  The  same.     Toxicite  du  blanc  d'oeuf;  1'influence  des  saisons  sur  la 

sensibilite  de  Porganisme  a  1'intoxication  azotee.     C.  r.  166,  919, 
1918. 


LITERATURE    TO    THE    TEXT  409 

195.  DAVIS,  W.  A.:  The  phosphate  depletion  of  the  soils  of  Bihar;  its  effect 

on  the  quality  and  yield  of  crops  and  the  contingent  risks  of  malnu- 
trition and  endemic  diseases  in  cattle  and  man.  Agr.  J.  India 
(Spec.  Ind.  Sci.  Congr.  No.)  77,  1917. 

196.  SCHOTTELIUS:  Arch.  f.  Hyg.  34,  1899;  42,  1902;  47,  1908. 

197.  BELONOWSKY:  Sur  1'alimentation  sterilisee.     Z.  f.  Bakt.  44,  322,  1907. 

198.  BOGDANOW,  E.  A.:  Ueber  das  Ziichten  der  Larven  der  gewohnlichen 

Hausfliege  (Calliphora  vomitoria)  in  sterilisierten  Nahrmitteln. 
Arch.  f.  ges.  Physiol.  113,  97,  1906;  Arch.  Physiol.  Suppl.  173,  1908. 

199.  DELCOURT,   A.   AND  GUYENOT,  E. :  Bull.  Sci.  France  et  Belg.  14,  249, 

1911;  Guyenot:  C.  r.  soc.  biol.  65,  Part  i,  97,  178,  223,  270,  1914; 
Part  i,  66,  483,  548,  1914;  Recherches  experimentales  sur  la  vie 
aseptique  et  le  developement  d'un  organisme  enfonctiondumillieu. 
These  Doct.  Sci.  Paris,  1917. 

200.  WOLLMANN,  N. :  Sur  la  vie  sans  microbes.     Ann.  Inst.  Past.  25,  79,  1911; 

La  vie  aseptique.  Bull.  Inst.  Past.  15  et  30  d<§c.  1914;  Elevage 
aseptique  des  larves  de  la  mouche  a  viande  (Calliphora  vomitoria) 
sue  millieu  sterilise  a  haute  temperature.  C.  r.  soc.  biol.  31  mai 
1919;  Richet,  Ch. :  L'alimentation  avec  les  aliments  ste"rilise"s  remarque 
a  propos  de  la  note  de  M.  Wollmann.  C.  r.  soc.  biol.  7  juin,  1919. 

201.  METCHNIKOFF,  MME.  :  cited  by  Cohendy.     Ann.  Inst.  Past.  27,  106,1912. 

202.  MORO:  Das  Leben  ohne  Bakterien.     J.  f.  Kinderh.  62,  H.  5,  1905. 

203.  COHENDY:  Experiences  sur  la  vie  sans  microbes.     Ann.  Inst.  Past.  26, 

106,  1912;  II.  Experiences  sur  la  vie  en  cultures  pures  succe"dant  a 
la  vie  sans  microbes.  C.  r.  154,  670,  1912. 

204.  The  same.     La  vie  aseptique.     C.  r.  158,  1283,  1914. 

205.  SCHOTTELIUS:  Die  Bedeutung  der  Darmbakterien  fur  die  Ernahrung, 

IV.  Arch.  f.  Hyg.  79,  1913. 

206.  KUSTER:  Leben  onhe  Bakterien.     Arb.  a.  d.  Kais.     Gesundheitamte. 

48,  1,  1914. 

207.  CHARRIN  AND  GUILLEMONAT:  cited  by  Weill-Mouriquand.    Maladies 

par  carence.     Bailliere,  Paris,  1919. 

208.  KIANIZIN,  J. :  Effect  on  higher  animals  of  the  sterilization  of  the  inhab- 

ited medium,  the  air  and  the  food.     J.  Physiol.  50,  391,  1916. 

209.  NUTTAL  AND  TniERFELDER:  cited  by  Kiister.     Die  keimfreie  Ziichtung 

von  Saugetieren.  Abderhaldens  H.  d.  Biochem.  Arbeitsm.  8,  311, 
1915. 

210.  WEILL,  E.  AND  MOURIQUAND,  G. :  C.  r.  soc.  biol.  81,  1253,  1918. 

211.  SCHAEFFER,   G. :  Conception  de  la  carence  dans  Interpretation  des 

resultats  obtenus  dans  les  travaux  sur  1'alimentation  artificielle  et 
la  vie  aseptique.  Ibid.  82,  2,  1919. 

212.  PORTIER,  PAUL.  :  Les  symbiotes.    Masson  &  Cie,  Paris,  1918. 

213.  BIERRY,  H.  AND  PORTIER,  P.:  Vitamines  et  symbiotes.     C.  r.  166,  963, 

1918;  Action  des  symbiotes  sur  les  constituants  des  graisses.  C.  r. 
166,  1055,  1918;  L'importance  de  la  function  cetonique  dans  la 
nutrition,  sa  creation  par  les  symbiotes.  Ibid.  167,  94,  1918. 

214.  DELAGE,  YVES:  Ibid.  166,  966,  1918. 


410  THE   VITAMINES 

215.  LUMIERE,  AUGUSTE:  Le  mythe  des  symbiotes.    Masson,  Paris,  1919. 

216.  RANG,  A.:  Bioenergesis  et  la  the"orie  des  symbiotes  de  Paul  Portier. 

L'ind.  chim.  6,  136,  1919. 

217.  PORTIER,  P.  AND  RANDOIN,  LUCIE  :  La  production  des  vitamines  dans 

les  intestins  des  lapins,  relevant  une  nourriture  sterilised  £  haute 
temperature.  C.  r.  170,  478,  1920. 

218.  LUND:  Am.  J.  Physiol.  47,  167,  1918. 

219.  CALKINS,  G.  W.  AND  EDDY,  W.  H. :  The  action  of  pancreatic  vitamine 

upon  the  metabolic  activity  of  Paramecium.  Proc.  Soc.  Exp. 
Biol.  Med.  14,  162,  1917. 

220.  FLATHER,  MARY  DRUSILLA:  Effects  of  polished  and  of  unpolished  rice 

upon  the  metabolic  activity  of  Paramecium.  Biol.  Bull.  36,  54, 
1919. 

221.  CHAMBERS,  MARY  H. :  Effect  of  some  food  hormones  and   glandular 

products  on  the  rate  of  growth  of  Paramecium  caudatum.  Ibid. 
36,  82,  1919. 

222.  PETERS,  R.  A. :  Nutrition  of  the  protozoa.    The  growth  of  Paramecium 

in  sterile  culture  medium  (Prelim.  Comm.).  Proc.  Physiol.  Soc. 
Febr.  21,  1920,  J.  Physiol.  53,  CVIII,  1920;  54,  L,  1920;  The  sub- 
stances needed  for  the  growth  of  a  pure  culture  of  Colpidium 
colpoda.  J.  Physiol.  55,  1,  1921. 

223.  GOLDFARB,  A.  J. :  A  study  of  the  influence  of  lecithin  on  growth.     Proc. 

Soc.  Exp.  Biol.  Med.  4,  159,  1907. 

224.  The  same.    Does  lecithin  influence  growth?    Arch.  Entw.  Mech.  29, 

255,  1910. 

225.  CARREL,  A.:  Technique  for  cultivating  a  large  quantity  of  tissue.     J. 

Exp.  Med.  15,  393,  1912. 

226.  The  same.    On  the  permanent  life  of  tissues  outside  of  the  organism. 

Ibid.  15,  516,  1912. 

227.  The  same.    Handb.  der  biochem.  Arbeitsm.  6,  519,  1912. 

228.  CARREL  AND  BURROWS  :  An  addition  to  the  technique  of  the  cultivation 

of  tissues  in  vitro.    J.  Exp.  Med.  14,  244,  1911. 

229.  CARREL,  A.:  Artificial  activation  of  the  growth  in  vitro  of  connective 

tissue.     Ibid.  17,  14. 

230.  WALTON,  A.  J. :  The  influence  of  internal  secretion  glands  on  growth. 

Ibid.  20,  554,  1914. 

231.  LOEB,  JACQUES:  The  simplest  constituent  required  for  growth  and  the 

completion  of  the  life  cycle  in  an  insect.    Science  41,  169,  1915. 

232.  The  same.     The  salts  required    for  the  development   of  insects.     J. 

Biol.  Chem.  23,  431,  1915. 

"  233.  LOEB,  JACQUES  AND  NORTHROP,  J.  H. :   Nutrition    and  evolution.     J. 
Biol.  Chem.  27,  309,  1916. 

234.  The  same.     The  influence  of  food  and  temperature  upon  the  duration 

of  life.     Ibid.  32,  103,  1917. 

235.  NORTHROP,  JOHN  H. :  The  effect  of  prolongation  of  the  period  of  growth 

on  the  total  duration  of  life.     Ibid.  32,  123,  1917. 

236.  The  same.     The  role  of  yeast  in  the  nutrition  of  an  insect  (Drosophils) . 

Ibid.  30,  181,  1917. 


LITERATURE    TO    THE    TEXT  411 

237.  GUYENOT,  EMILE:  Bull.  biol.  de  France  et  de  la  Belg.  51,  14  aout  et  15 

d6c.  1917. 

238.  BAUMBERGER,  J.  PERCY:  A  nutritional  study  of  insects,  with  special 

reference   to  microorganisms   and  the  substrate.     J.   Exp.   Zool. 
28,  1,  1919. 

239.  MORGULIS,  SERGIUS:  Studies  on  the  nutrition  of  the  fish.    Experiments 

on  brook  trout.     J.  Biol.  Chem.  36,  391,  1918. 

240.  ALMY,  L.  H.  AND  ROBINSON,  R.  K. :  Toxic  action  of  ingested  linseed  oil 

meal  on  trout.     J.  Biol.  Chem.  43,  97,  1920. 

241.  EMMETT,  A.  D.  AND  ALLEN,  FLOYD  P. :  Nutritional  studies  on  the  growth 

of  frog  larvae  (Rana  pipiens).     Ibid.  38,  325,  1919. 

242.  EMMETT,  ALLEN  AND  STURTEVANT,  MARGUERITE  :  Relation  of  vitamines 

and  iodine  to  the  size  and  development  of  the  tadpoles.     Proc. 
Soc.  Amer.  Biol.  Chem.     J.  Biol.  Chem.  41,  LIV,  1920. 

243.  HARDEN,  ARTHUR  AND  ZILVA,   SOLOMON   SYLVESTER:    Dietetic  experi- 

ments with  frogs.     Biochem.  J.  14,  263,  1920. 

244.  FUNK,  CASIMIR:  Studien  liber  das  Wachstum.     I.  Das  Wachstum  auf 

vitaminhaltiger  und  vitaminfreier  Nahrung.    Ztschr.    f.    physiol. 
Chem.  86,  352,  1913. 

245.  FUNK,  CASIMIR  AND  MACALLUM,  A.  B. :  Die  chemischen  Determinanten 

des  Wachstums.     Ibid.  92,  13,  1914. 

246.  FUNK,  CASIMIR:  Studies  on  growth:  the  influence  of  diet  on  growth, 

normal  and  malignant.     Lane.  186,  89,  1914. 

247.  DRUMMOND,  J.  C. :  Observations  upon  the  growth  of  young  chickens 

under  laboratory  conditions.     Biochem.  J.  10,  77,  1916. 

248.  FUNK,  CASIMIR:  J.  Biol  Chem.  27,  1,  1916  (Footnote  p.  4). 

249.  BUCKNER,  G.  D.,  NOLLAU,  E.  H.  AND  KASTLE,  J.  H. :  Feeding  of  young 

chicks  on  grain  mixture  of  high  and  low  lysine  content.    Am.  J. 
Physiol.  39,  162,  1915. 

250.  OSBORNE  AND  MENDEL:  The  effect  of  the  amino-acid  content  of  the  diet 

on  the  growth  of  chicken.     J.  Biol.  Chem.  26,  293,  1916. 

251.  The  same.     The  growth  of  chickens  in  confinement.     Ibid.   33,   433, 

1918;  J.  Pharmac.  11,  170,  1918. 

252.  HART,  E.  B.,  HALPIN,  J.  C.  AND  STEENBOCK,  H. :  Use  of  synthetic  diets 

in  the  growth  of  baby  chicks.     A  study  of  leg  weakness  in  chickens. 
J.  Biol.  Chem.  43,  421,  1920. 

253.  PALMER,  LEROY  S. :  The  physiological  relation  of  plant  carotiniods  to 

the  carotinoids  of  the  cow,  horse,  sheep,  goat,  pig  and  hen.     Ibid. 
27,  27,  1916. 

254.  PALMER  AND  KEMPSTER,  HARRY  L. :  Relation  of   plant  carotinoids  to 

growth,  fecundity,  and  reproduction  of  fowls.     Ibid.  39,  299,  313, 
333,  1919. 

255.  HART,  E.  B.,  HALPIN,  J.  G.  AND  McCoLLUM,  E.  V.:  Ibid.  29,  57,  1917. 

256.  HART,  HALPIN  AND  STEENBOCK:  The  behavior  of  chickens  restricted  to 

the  wheat  and  maize  kernel.     II.  Ibid.  31,  415,  1917. 

257.  HARNEY,  R.  N. :  Egg-producing  values  of  some  Texas  feeding  stuffs. 

Texas  Agr.  Exp.  Sta.  Bull.  220,  11,  1917. 


412  THE   VITAMINES 

258.  KAUPP,  B.  F. :  Mineral  content  of  Southern  poultry  feeds  and  mineral 

requirement  of  growing  fowls.     J.  Agr.  Res.  14,  125,  1918. 

259.  McCoLLUM  AND  DAVIS,  MARGUERITE:  The  nature  of  the  dietary  defi- 

ciency in  rice.     J.  Biol.  Chem.  23,  181,  1915. 

260.  HOULBERT:  Vitamines  et  la  croissance.     Paris  me"d.  9,  473,  1919. 

261.  HILL,  L.  AND  FLACK,  M. :  Brit.  Med.  J.  Sept.  16th,  311,  1911. 

262.  OHLER,  RICHARD  W. :  Experimental  polyneuritis — Effects  of  exclusive 

diet  of  wheat  flour,  in  form  of  ordinary  bread,  in  fowls.  J.  Med. 
Res.  31,  239,  1914. 

263.  WEILL,  E.  AND  MOURIQUAND,  G. :  Troubles  provoques  par  une  alimen- 

tation exclusive.     Ann.  de  me'd.  et  chir.  infant.  18,  mai  15,  1914. 

264.  WELLMAN,  BASS  AND  EUSTIS:  Investigations  of  Louisiana  rice,    with 

reference  to  the  etiology  of  beriberi.  New  Orl.  Med  Surg.  J.  65, 
191,  1912.  Wellmann:  South.  Med.  J.  6,  516,  1913.. 

265.  WELLMAN,   C.,  AND  BASS,  C.   C. :    Polyneuritis    gallinarum     caused     by 

different  foodstuffs.    Am.  J.  Trop.  Dis.  and  Prev.  Med.  1,  129,  1913. 

266.  VEDDER  AND  CLARK:  A  study  of  polyneuritis  gallinarum.    A  fifth  contri- 

bution to  the  etiology  of  beriberi.     Philipp.  J.  Sci.  7  B,  423,  1912. 

267.  SEGAWA:  Ingestion  of  polished  rice  by  chickens  and  pigeons.     Mitt. 

med.  Ges.  Tokyo  27  No  7,  1914;  Virch.  Arch.  215,  404,  1914.. 

268.  TASAWA:  Experimentelle  Polyneuritis,  besonders  bei  Vogeln,  im  Ver- 

gleich  zur  Beriberi  des  Menschen.  Z.  f.  exp.  Path.  Ther.  17,  27, 
1915. 

269.  POL,  HULSHOFF:  Geneesk.  Tijdsr.  v.  Ned.  Indie.  52,  11,  245,  1912. 

270.  WILLIAMS,  R.  R.  AND  JOHNSTON,  J.  A. :   Miscellaneous    notes  and  com- 

ments on  beriberi.     Philipp.  J.  Sci.  10B,  337,  1915. 

271.  VOEGTLIN,  CARL  AND  MYERS,  C.  N. :  Distribution  of   the  antineuritic 

vitamine  in  the  wheat  and  corn  kernel.  A  contribution  to  the 
biology  of  the  antineuritic  vitamine.  Am.  J.  Physiol.  48,  504,  1919. 

272.  The  same.    Growth-promoting  properties  of  foods  derived  from  corn 

and  wheat.    U.  S.  Public  Health  Serv.  Repr.  471,  23,  1918. 

273.  FUNK,  CASIMIR:  Is  polished  rice  plus  vitamine  a  complete  food?    J. 

Physiol.  48,  228,  1914. 

274.  STEPP,  WILHELM:  Zur  Frage  der  synthetischen  Fahigkeiten  des  Tier- 

korpers.  Z.  f.  Biol.  66,  300,  1916. 

275.  FUNK,  CASIMIR:  Studien  iiber  Beriberi  XI.  Die  Rolle  der  Vitamine 

beim  Kohlenhydratstoffwechsel.  Ztschr.  f.  physiol.  Chem.  89, 
378,  1914. 

276.  DUTCHER,  R.  ADAMS:  Vitamine  Studies  IV.    Antineuritic  properties  of 

certain  physiological  stimulants.  Prelim.  Paper.  J.  Biol.  Chem. 
39,  63,  1919. 

277.  COOPER,  E.  A.  AND  FUNK,  CASIMIR:  Experiments  on  causation  of  beri- 

beri.    Lane.  Nov.  4th,  p.  1266,  1911. 

278.  THEILER,  A.  GREEN,  H.  H.,  AND  VILJOEN,  P.    R. :  Contribution   to 

the  study  of  deficiency  diseases,  with  special  reference  to  the 
lamziekte  problem  im  South  Africa.  3rd  and  4th  Director  Veter. 
Res.  Rep.  Dept.  Agr.  Union  S.  Africa  9,  1915. 


LITERATURE    TO    THE    TEXT  413 

279.  LUMIERE,  A. :  Bull  de  1'acad.  de  meU  83,  96,  1920. 

280.  MERKLEN,    P. :  Modification   of  diet   saves  ducklings  from   epidemic 

diseases.     Bull,  de  la  soc.  de  p6d.  16,  April,  1914. 

281.  KiiLZ,  L.:  Ueber  Beriberi  derEnten.  Arch.  Schiffs-  und  Tropenh.  16,  163, 

1912. 

282.  FUJITANI,  J. :  Beitrage  zur  aetiologischen  Kenntnis  der  bei  Reisfiitterung 

auftretenden  Krankheit  der  Vogel.     Mitt.   Berib.   Stud.     Komm. 
Tokyo.  306,  1911. 

283.  TOYAMA,  C. :  Ueber  eine  kakkeahnliche  Krankheit  der  Vogel.      Ibid. 

274,  1911. 

284.  FINK,  G.  L. :  Beriberi  und  weisser  Reis,  cin  Versuch  mit  Papageien.     J. 

Trop.  Med.  Hyg.  15,  VIII,  1910;  Arch.  Schiffs  u.  Tropenhyg.   15, 
270,  1911. 

284a.  JANSEN,  B.  C.  P. :  Vitamine  content  of  extract  of  rice  bran.    Medel. 
v.  I.  Burg.  Dienst.  Ned.  Indie.  No.  1,  23,  1920. 

285.  KIMURA,  ONARI:  Degeneration  and  regeneration  in  peripheral  nerves. 

Mitt.  Path.  Inst.  Univers.  Sendai,  Japan,  March  24th,  1919. 

286.  SCHNYDER,  K. :  Patkologisch-anatomische  Untersuchungen  bei  experi- 

mentellen  Beriberi  (Reispolyneuritis)  Arch.  f.  Verdauungskrankh. 
20,  147,  1914. 

287.  WEILL  AND  MOURIQUAND:  Neurologic  expe"rimentale.     Les   paralysies 

par  carence.  Soc.  de  neurol.  Mai,  1917. 

288.  KATO,  G.  AND  SHIZUME,  S. :  Physiologic  and  pathologic  examination  of 

nerves    and   muscles   of   domestic    fowl    suffering   from   so-called 
"polished  rice  disease".     Japan  Med.  World.  Nov.  23,  No.  310,  1919. 

289.  PAGUCHI,  K. :  Ibid.  Dec.  14th,  No.  313,  1919. 

290.  McCARRisoN,  R. :  Genesis  of  edema  in  beriberi.     Proc.  R.  Soc.  91  B,  103, 

1920;  Pathogenesis  of  deficiency  diseases  II.     Effects  of  deprivation 
of  "B"  accessory  food  factor.     Ind.  J.  Med.  Res.  6,  550,  1919. 

291.  FUNK,  CASIMIR  AND  DOUGLAS,  MACKENZIE:  Studies  on   beriberi  VIII. 

The  relationship  of  beriberi  to  glands  of  internal  secretion.     J. 
Physiol.  47,  475,  1914. 

292.  WILLIAMS,  R.  R.  AND  CROWELL,  B.   C. :  Thymus    gland  in   beriberi. 

Philipp.  J.  Sci.  10  B,  121,  1915. 

293.  MCCARRISON,  R. :  Involution  of  thymus  in  birds.     Ind.  J.   Med.  Res. 

6,  557,  1919. 

294.  DOUGLAS,  MACKENZIE:  The  histology  of  the  thyroid  in  animals  fed  on 

various  diets.     J.  Path.  Bact.  19,  341,  1915. 

295.  MCCARRISON,   R. :  Pathogenesis  of  deficiency  diseases.     X.  Effect  of 

some  food  deficiencies  and  excesses  in  thyroid  gland.     Ind.  J.  Med. 
Res.  7.  633,  1920. 

296.  MCCARRISON:  Ibid.  5,  2755,  1919. 

297.  MCCARRISON:  Ibid.  2,  369,  1914. 

298.  MCCARRISON:  Pathogenesis  of  deficiency  diseases.     V.  Histopathology. 

Ibid.  7,  269,  1919. 

299.  MCCARRISON:  The  influence  of  deficiency  of  accessory  food  factors  on 

the  intestine.     Brit.  Med.  J.  July  12th,  1919. 


414  THE   VITAMINES 

300.  MCCARRISON:    The    pathogenesis    of    deficiency    diseases.     III.   The 

influence  of  dietaries  deficient  in  accessory  food  factors  on  the 
intestine.  Ind.  J.  Med.  Res.  7,  167,  1919. 

301.  FUNK,  CASIMIR:  The  effect  of  a  diet  of  polished  rice  on  the  nitrogen  and 

phosphorus  of  the  brain.     J.  Physiol.  44,  51,  1912. 

302.  WIELAND,  HERMANN:  Analytische  Untersuchungen  iiber  den   P-gehalt 

der  ernahrungskranken  Tieren.  Arch.  exp.  Path.  Pharm.  69,  93, 
1912. 

303.  KOCH,  MATHILDE  AND  VOEGTLIN,  CARL:  Chemical  changes  in  the  central 

nervous  system  as  a  result  of  restricted  vegetable  diet.  U.  S.  Publ. 
Health  Serv.  Hyg.  Lab.  Bull.  103,  1916. 

304.  FUNK,  CASIMIR  AND  v.   SCHONBORN:  The  influence    of    vitamine-free 

diet  on  the  carbohydrate  metabolism.     J.  Physiol.  48,  328,  1914. 

305.  FUGI,  S.:  Diastase  in  blood  of  chickens  fed  on  polished  rice.     Jikwa 

Zasshi  No.  203,  36,  1917. 

305a.  FINDLAY,    GEORGE  MARSHALL:     Glyoxylase    in   beriberi.    Biochem. 
J.  15,  104,  1921. 

306.  FUNK,  CASIMIR:  Ueber  die  physiologische  Bedeutung  gewisser  unbisher 

bekannter  Nahrungsbestandteile,  der  Vitamine.  Erg.  d.  Physiol. 
13,  125,  1913. 

307.  BENEDICT,  STANLEY  R. :  Uric  acid  in  its  relation  to  metabolism.     J. 

Lab.  Clin.  Med.  2,  No.  1,  1916. 

308.  HUNTER,  ANDREW  AND  WARD,  F.  W. :  Purine  metabolism   in  various 

representative  mammals.  Trans.  R.  Soc.  Canada.  13,  Section  IV, 
7,  1919. 

309.  MCCOLLUM,  E.  V.,  SIMMONDS,  N.  AND  PITZ,  W. :  The  effects  of  feeding 

the  proteins  of  the  wheat  kernel  at  different  planes  of  intake.  J. 
Biol.  Chem.  28,  211,  1916. 

310.  JACKSON,  C.  M. :  Biol.  Bull.  23,  171,  1917. 

311.  MACALLUM,  ARCHIBALD  BRUCE:  The  relation  of  vitamines  to  the  growth 

of  young  animals.     Trans.  R.  Can.  Inst.  p.  175,  1919. 

312.  FERRY,  EDNA  L. :  Nutrition  experiments  with  rats.     J.  Lab.  Clin.  Med. 

5,  735,  1910. 

313.  DRUMMOND,  J.  C. :  The  growth  of  rats  upon  artificial  diet  containing 

lactose.     Biochem.  J.  10,  89,  1916. 

314.  OSBORNE  AND  MENDEL:  J.  Biol.  Chem.  34,  309,  1918. 

315.  McCoLLUM  AND  SIMMONDS  :  A  study  of  the  dietary  essential  water-soluble 

B,  in  relation  to  its  solubility  and  stability  towards  reagents. 
Ibid.  33,  55,  1918. 

316.  OSBORNE  AND  MENDEL:  Ibid.  18,  95,  1914;  The  suppression  of  growth 

and  the  process  of  growth.     Ibid.  23,  439,  1915. 

317.  JACKSON,  C.  M.  AND  STEWART,  C.  A. :  The  effects  of  underfeeding  and 

refeeding  upon  the  growth  of  various  systems  and  organs  of  the 
body.  Minn.  Med.  Nov.,  1918;  J.  of  Zool.  30,  97,  1920. 

318.  BRUNING,  H. :  Experimentelle  Studien  iiber  die  Entwickelung   neuge- 

borener  Tiere  bei  langerdauernden  Trennung  von  der  saugenden 
Mutter  und  nachheriger  verschiedenartiger  klinstlichen  Ernahrung. 
J.  f.  Kinderh.  80,  65,  1914. 


LITERATURE   TO   THE   TEXT  415 

319.  ARON,  HANS:  Untersuchungen  iiber  die  Beeinflussung  des  Wachsturas 

durch  Ernahrung.     Berl.  klin.  W.  51,  972,  1914. 

320.  SLONACKER,  J.  R. :  Leland  Stanford,  Jun.  Univers.  Public.  1912. 

321.  OSBORNE  AND  MENDEL:  The  choice  between  adequate  and  inadequate 

diets,  as  made  by  rats.     J.  Biol  Chem.  35,  19,  1918. 

322.  SURE,  BARNETT:  The  nutritive  value  of  lactalbumin;  cystine  and  tyro- 

sine  as  growth-limiting  factors  in  that  protein.     Ibid.  43,  457,  1920. 

323.  SUZUKI,  UMETARO,  OKUDA,  YUZURU,  MATSUYAMA,  YOSHIHIKO,  OKIMOTO, 

TAMAO,  KATAKURA,  KEI  AND  IWATA,  MOTOE:  Nutritive  value  of 
various  proteins  and  fats.     J.  Tokyo  Chem.  Soc.  41,  381,  1920. 

324.  FUNK,  CASIMIR:  The  preparation  from  yeast  and  certain  foodstuffs  of 

the  substance,  the  deficiency  of  which  occasions  polyneuritis  in 
birds.     J.  Physiol.  45,  75,  1912. 

325.  The  same.     The  study  of  certain  dietary  conditions  bearing  on  the 

problem  of  growth  in  rats.     J.  Biol.  Chem.  27,  1,  1916. 

326.  OSBORNE  AND  MENDEL:  Milk  as  a  source  of  water-soluble  vitamine. 

Ibid.  34,  537,  1918;  II  41,  515,  1920. 

327.  STEPP,  W. :  Weitere  Untersuchungen  iiber  die  Unentbehrlichkeit  der 

Lipoide   fur   das   Leben.     Ueber   die   Hitzezerstorbarkeit   lebens- 
wichtiger  Lipoide  der  Nahrung.     Z.  f.  Biol.  59,  336,  1912. 

328.  LANDER,  PERCY  E. :  On  the  cholesterol  content  of  the  tissues  of  growing 

rats  when  under  various  diets.     Biochem.  J.  9,  78,  1915. 

329.  ARON,  H. :   Die   Bedeutung   von   Extraktstoffen   fur   die   Ernahrung. 

Monatsh.  Kinderh.  13,  359,  1915. 

330.  FUNK,  CASIMIR  AND  MACALLUM,  A.  B.,   JR.:  Studies  on  growth.    III. 

The  comparative  value  of  lard  and  butter  fat  in  growth.     J.  Biol. 
Chem.  27,  51,  1916. 

331.  FUNK,  CASIMIR  AND  DUBIN,  HARRY  E. :  The  vitamine  requirements  of 

the  rat  on  diets  rich  in  protein,  carbohydrate  and  fat  respectively. 
Science,  N.  S.,  52,  447,  1920. 

332.  DESGREZ,  A.  AND  BIERRY,  H. :  L'equilibre   d'azote    et  1'absence  des 

vitamines.     C.  r.  170,  1209,  1920. 

333.  Report  of  Medical  Research  Committee  on  the  present  state  of  knowledge 

concerning  accessory  food  factors  (vitamines).     No.  38,  1919. 

334.  VOEGTLIN,  Carl  AND  LAKE,  G.  C. :  Experimental  mammalian  polyneuritis 

produced  by  a  deficient  diet.    Am.  J.  Physiol.  48,  558.  1919. 

335.  OSBORNE,  T.  B.:  The  water-soluble  vitamine.     N.  Y.  State  Med.    J. 

20,  217,  1920. 

336.  OSBORNE  AND  MENDEL.  The  role  of  vitamines  in  the  diet.     J.  Biol. 

Chem.  31,  149,  1917. 

337.  DRUMMOND,   J.   C. :  A  study  of  the  water-soluble  accessory  growth- 

promoting  substance.     II.    Its  influence  upon  the  nutrition  and 
nitrogen  metabolism  of  the  rat.     Biochem.  J.  12,  25,  1918. 

338.  OSBORNE  AND  MENDEL:  The  nutritive  value  of  yeast  protein.     J.  Biol. 

Chem.  38,  223,  1919. 

339.  EMMETT,  A.  D.  AND  ALLEN,  F.  P. :  Pathogenesis  due  to  vitamine  defi- 

ciency in  the  rat.     Proc.  Soc.  Amer.  Biol.  Chem.  XIV,  J.  Biol. 
Chem.  41,  LIII,  1920. 


416  THE    VITAMINES 

339a.  TSULI,  K.:    Effect  of  diet  on   thyroid    functioning.     Acta    Scholae 
Med.  Kyoto,  3,  713,  1920. 

340.  ARON,  HANS:  Berl.  klin.  W.  No.  23,  546,  1918. 

340a.  MORGULIS,  S.  AND  GIES,  W.  J. .  Calcium  content  of  bones  and  teeth 

from  normal  and  thymectomized  albino  rats.     J.  Exp.  Med.  20, 

499,  1914. 
340b.  MAT-TILL,  H.  A.:    Influence  of  fasting  and  vitamine  B  deprivation  on 

the  non-protein  nitrogen  of  rat's  blood.     Science,  54,  176,  1921. 
340c.  BRUNING,  H. :  Untersuchungen  iiber  das  Wachstum  von  Tieren  jenseits 

der  Sanglingsperiode  by  verschiedenartiger  kiinstlichen  Ernahrung. 

J.  f.  Kinderh.  79,  305,  1914. 

341.  MACALLUM,  ARCHIBALD  BRUCE:  The  relationship  of  vitamines  to  animal 

growth.    Amer.  Med.  11,  782,  1916. 

341a.  HOPKINS,  F.  G.:    Note  on  vitamine  content  of  milk.     Biochem  J. 
14,  721,  1920. 

342.  FREISE,  E. :  Nahrungsbestandteile  die  das  Wachstum  erregen.     J.    f. 

Kinderh.  91,  79,  1920. 

342a.  FUNK,   CASIMIR  AND  DUBIN,  HARRY  E.:    Vitamines    of    autolyzed 
yeast  in  nutrition.    Soc.  Exp.  Biol.  Med.    Oct.  19,  1921. 

343.  ROBERTSON,  T.  BRAILSFORD:  The  normal  growth  of  the  white  mouse. 

J.  Biol.  Chem.  24,  363,  1916. 

344.  THOMPSON,  HEEEN  B.,  AND  MENDEL,  L.  B. :  Alternating  growth  and 

suppression  of  growth  in  the  albino  mouse,  with  special  reference 
to  the  economy  of  food  consumption.     Am.  J.  Physiol.  45,  431,  1918. 

345.  MENDEL  AND  JUDSON,  S.  E. :  Some  interrelations  between  diet,  growth 

and  chemical  composition  of  the  body.     Proc.  Nat.  Acad.  Sci.  2, 
692,  1916. 

346.  SCHMIDT,  M.  B.:  Eisenstoffwechsel  und  Blutbildung.  Mitt.  d.  Physik. 

Med.  Ges.  Wiirzburg.  Ref.  D.  med.  W.  40,  205,  1914;  Verh.  d.  deutsch. 
path.  Ges.  17  Tagung,  156,  1914. 

347.  MORPURGO,  B.  AND  SATTA,  G. :  Sugli  scambi  de  sostanze  nutrienti  fra 

topi  in  parabiosi.     Arch,  di  fisiol.  11,  360,  1914. 

348.  STEPP,  W. :  Untersuchungen  liber  die  Unentbehrlichkeit  der  Lipoide 

fur  das  Leben.     Z.  f .  Biol.  62,  405,  1913. 

349.  ROHL,    W. :   Experimented   Untersuchungen   tiber    den   Aufbau    von 

Lipoiden   im   Tierkorper.     Verh.    des  29en  deutsch.  Kongr.  f .    inn. 
Med.  Wiesbaden,  1912. 

350.  STEPP,   W. :  Ueber  lipoidfreie  Ernahrung  und  ihre  Beziehungen  zur 

Beriberi  und  Skorbut.     D.  med.  W.  40,  892,  1914. 

351.  STEPP:  Lipoid  Hunger  und  Beriberi.     Z.  f.  Biol.  66,  339,  1916. 

352.  STEPP:  Die  Unentbehtlichkeit  der  Lipoide  fur  das  Leben.     Ibid.  66, 

365,  1916. 

353.  DEZANI,  SERAFINO:  Untersuchungen  iiber  die  Genese  des  Cholesterins. 

Arch.  farm.  17,  4,  1914. 

354.  DEZANI  :  Ricerche  sulla  nutrizione  alipoidea.     Biochimica.  4,  475,  1914. 

355.  ROHMANN,  F. :  Ueber  die  Ernahrung  von  Mausen  mit  einer  aus  einfachen 

Nahrungstoffen  zusammengesetzter  Nahrung.     Bio.  Z.  64,  30,  1914. 


LITERATURE   TO   THE   TEXT  417 

356.  WHEELER,  RUTH:  Feeding  experiments  with  mice.     J.  Exp.  Zool,  15, 

209,  1913. 

357.  MACARTHUR,  C.  G.  AND  LUCKETT,  C.  L. :  Lipins  in  nutrition.     J.  Biol. 

Chem.  30,  161,  1915. 

358.  MITCHELL,  H.  H.  AND  NELSON,  B.  A. :  The  preparation  of  protein-free 

milk.     J.  Biol.  Chem.  23,  459,  1915. 
358a.  HUME,  E.  M.:    Comparison  of  the  growth-promoting  properties  for 

guinea  pigs  of  certain  diets  consisting  of  natural  foodstuffs.    Bio- 

chem.  J.  15,30,  1921. 
358b.  TOZER,  F.  M. :    Effect  on  guinea  pigs  of  deprivation  of  vitamine  A  and 

of  antiscorbutic  factor.    J.  Path.  Bact.  24,  306,  1921 . 

359.  SMITH,  THEOBALD  :  Bacilli  in  swine  disease.     Bur.  of  Anim.  Ind.  Wash. 

172,  1895-6. 

360.  HOLST,  AXEL  AND  FROLICH,  TH.  :  Ueber   experimentellen   Skorbut.  Z. 

f.  Hyg.  u.  Infektionsk.  72,  1,  1912. 

361.  JACKSON  AND  MOORE:  J.  Inf.  Dis.  19,  510,  1916. 

362.  JACKSON  AND  MOODY:  Ibid.  19,  511,  1916. 

363.  MOORE,  J.  J.  AND  JACKSON,  L. :  Experimental  scurvy  produced  in  guinea 

pigs  by  milk  and  milk  products.     J.  A.  M.  A.  67,  1931,  1916. 

364.  GIVENS,  MAURICE  H.  AND  HOFFMAN,   GEORGE  L. :  Preliminary  obser- 

vations on  the  relation  of  bacteria  to  experimental  scurvy  in  guinea 
pigs.  Proc.  Soc.  Amer.  Biol  Chem.  XIV.  J/  Biol.  Chem.  41, 
XXXIII,  1920. 

365.  PITZ,  W. :  Studies  on  experimental  scurvy.     J.  Biol.  Chem.  33,  471, 1918; 

36,  439,  1918. 

366.  TORREY,  J.  C.  AND  HESS,  A.  F. :  The  relation  of  the  intestinal  flora  to 

the  scurvy  of  guinea  pigs  and  of  infants.  Proc.  Soc.  Exp.  Biol. 
15,  74,  1918. 

367.  HARDEN  AND  ZILVA:  Note  on  the  etiology  of  scurvy  in  guinea  pigs. 

Biochem.  J.  12,  270,  1918. 

368.  FUNK,  CASIMIR:  Nature  of  the  disease  due  to  the  exclusive  diet  of  oats 

in  guinea  pigs  and  rabbits.     J.  Biol.  Chem.  25,  409,  1916. 

369.  HEIM,  P. :  Die  Bedeutung  der  Extrakstoffe  fur  die  Ernahrung.    Mon. 

f.  Kinderh.  13,  367,  1914-6;  13,  477,  1916. 

370.  RONDONI,  PIETRO  AND  MoNTAGNANi,  MARIO  :   Lesioni    istologiche  nel 

maidismo  nel  digiuno  e  nello  scorbuto  sperimentale.  H  Speriment. 
69,  659,  1915. 

371.  INGIER,  A.:  Ueber  experimentell  hervorgerufenen  Morbus  Barlow  an 

Foten  und  Neugeborenen.  Nord.  Med.  Ark.  48,  1,  1915;  Abt.  II; 
J.  Exp.  Med.  21,  525,  1915. 

372.  McCoLLUM,  SIMMONDS  AND  PITZ  :  The  nature  of  dietary  deficiencies 

of  the  oat  kernel.     J.  Biol.  Chem.  29,  341,  1917. 

373.  CHICK,  HARRIETTS  AND  HUME,  E.  MARGARET:  Trans.  Soc.  Trop.  Med.  & 

Hyg.  10,  152,  1917. 

374.  COHEN,  BARNETT  AND  MENDEL,  L.  B. :   Experimental   scurvy  of  the 

guinea  pig  in  relation  to  the  diet.     J.  Biol.  Chem.  35,  425,  1918. 

375.  HESS,  A.  F.  AND  UNGER,  L.  J. :  Experiments  on  scurvy  of  guinea  pig. 

Proc.  Soc.  Exp.  Biol.  Med.  15,  82,  1918;  J.  Biol.  Chem.  35,  479,  1918. 


418  THE    VITAMINES 

376.  CHICK,   HARRIETTS   AND  co-workers:   Cited  according    to  Med.    Res. 

Comm.  Rep.  No.  38.  p.  43,  1919. 

377.  HESS,  A.  F. :  The  antiscorbutic  vitamine.     N.  Y.  State  J.  of  Med.  20, 

209,  1920. 

378.  HESS  AND  UNGER.  Scorbutic  beading  of  ribs.   Amer.   J.   Dis.    Childr. 

19,  331,  1920;  Br.  M.  Assocn.  Meeting  June  29  Cambridge,  Br. 
Med.  J.  July  31,  154,  1920. 

379.  RONDONI,  P. :    Pathogenesis  of  deficiency  diseases  and  pellagra.   Br. 

Med.  J.  I,  542,  1919. 

380.  MCCARRISON,  R. :    Influence  af  scorbutic  diet  on  suprarenals.  Ind.  J. 

Med.  Res.  7,  188,  1919;  Br.  Med.  J.  Aug.  16th,  200,    1919. 

381.  LAMER,  V.  K.  AND  CAMPBELL,  L.  H. :  Changes  in  organ  weight  produced 

by  deficient  diet  in  antiscorbutic  vitamine.  Proc.  Soc.  Exp.  Biol. 
Med.  1920. 

382.  MCCARRISON,  R. :  Pathogenesis  of  deficiency  diseases.     VI.    Influence 

of  scorbutic  diet  on  bladder.     Ind.  J.  Med.  Res.  7,  279,  1919. 

383.  BAUMAN,  L.  AND  HOWARD,  C.  P. :  The  mineral  metabolism  of  experi- 

mental scurvy  in  the  guinea  pig.    Amer.  J.  Med.  Sci.  153,  650,  1917. 

384.  SMITH,  MILLARD  L.  AND  LEWIS,    HOWARD  B.:   Study  of  the  normal 

metabolism  of  the  guinea-pig.     J.  Amer.  Chem.  Soc.  39,  2231,  1917. 

385.  McCLENDON,  J.  F.,  COLE,  W.  C.  C.,  ENGSTRAND,  O.  AND  MIDDLEKAUFF, 

J.  E. :  The  effect  of  malt  and  malt  extracts  and  the  alkaline  reserve 
of  the  blood.  J.  Biol.  Chem.  40,  243,  1919. 

386.  LEWIS,  HOWARD  B.  AND  KARR,  WALTER  G. :  Changes  in  the  urea  content 

of  the  blood  and  tissues  of  guinea  pigs  maintained  on  an  exclusive 
oat  diet.  J.  Biol.  Chem.  28,  17,  1916. 

387.  KARR,  W.  G.  AND  LEWIS,  H.  B. :  The  phenol  excretion  of  guinea  pigs 

maintained  on  an  exclusive  oat  diet.  Amer.  J.  Physiol.  44,  586, 
1917. 

387a.  McCoLLUM   AND   PARSONS;   The   antiscorbutic    requirements   of    the 
prairie  dog.     J.  Biol.  Chem.  44,  603,  1920. 

388.  MORGEN,  A.  AND  BEGER,  C. :  Ueber  den  schadlichen,    auf  einer  Saure- 

vergiftung  zuriickfuhrenden  Einfluss  einer  ausschiesslichen  Ernah- 
rung.  H.  94,  324,  1915. 

389.  KURIJAMA,  S. :  J.  Biol.  Chem.  33,  215,  1918. 

390.  MCCLENDON,  J.  F.,  v.  MEYSENBUG,  L.,  ENGSTRAND,  O.  J.  AND  KING, 

FRANCIS:  Effect  of  diet  on  the  alcaline  reserve  of  the  blood.  Ibid. 
38,  539,  1919. 

391.  NELSON,  V.  E.  AND  LAMB,  A.  R. :  The  effect  of  vitamine  deficiency  in 

various  species  of  animals.  I.  The  production  of  xerophtalmia  in 
the  rabbit.  Amer.  J.  Physiol.  51,  530,  1920. 

392.  WEILL,   E.,  MOURIQUAND,   G.   AND  MICHEL,    P.:  Recherches    sur    les 

maladies  par  carence;  effets  compares  d'alimentation  exclusive  des 
chats,  avec  la  viande  crue,  congelee,  salee,  buillie  et  sterilisee.  C. 
r.  soc.  biol.  79,  189,  1916. 

393.  VOEGTLIN,  CARL  AND  LAKE,  G.  C. :  Experimental  mammalian  polyneu- 

ritis  produced  by  a  diet  deficient  in  antineutiric  "vitamine."  J. 
Pharmac.  11,  167,  1918;  Amer.  J.  Physiol.  48,  558,  1919. 


LITERATURE    TO    THE    TEXT  419 

393a.  MACKAY,  HELEN  MARION  MACPHERSON:    The  effect  on  kittens  of  a 
diet  deficient  in  animal  fat.    Biochem.  J.  15,  19,  1921. 

394.  BLAND-SUTTON  :    Cited  according  to  Cheadle  und  Poynton.     Allbutt's 

System  of  Medicine,  3,  85. 

395.  CAHN,  A.:  Die  Magenverdauimg  im  Chlorhunger.     Ztschr.  f.   Physiol. 

Chem.  10,  517,  1886. 

396.  TRAPPE,  K. :  Ueber  Saurebildung  im  Magen.     Diss.  Halle.  1892. 

397.  ROSEMANN,  R. :  Ueber  den  Chlorgehalt  des  tierischen  Korpers.     Pfl. 

Arch.  135,  177,  1910;  142,  208,  1911. 

398.  BONNIGER,  M. :   Die  Substituirung  des  Chlors  durch  Brom  im  Tier- 

korper.  Z.  f.  exp.  Path.  u.  Ther.  4,  414,  1907;  7,  556,  1911;  14,  452, 
1913. 

399.  PFLUGER:  Cited  according  to  Hofmeister.     Erg.  d.  Physiol.  16,  540,  1918. 

400.  HEUBNER,  WOFLGANG:  M.  med.  W.  2543,  1911;  Lipschiitz,  Alexander. 

Untersuchung  uber  den  Phosphorhaushalt  des  wachsenden  Hundes. 
Arch.  f.  exp.  Path.  Pharm.  62,  210,  1910;  Arch.  f.  ges.  Physiol. 
143,  91,  1911;  Durlach,  E.  Arch.  f.  exp.  Path.  Pharm.  71,  210,  1913; 
Heubner.  Ueber  den  P-gehalt  tierischer  Organe  nach  verschieden- 
artiger  Fattening.  Ibid.  78,  22,  1915. 

401.  SCHMORL,  C. :  Ueber  die  Beeinflussung  des  Knochenwachstums  bei  phos- 

phorarmer  Ernahrung.     Ibid.  73,  313,  1913. 

402.  MASSLOW:  Ueber  die  biologische  Bedetitung  des  Phosphors    fur  den 

wachsenden  Organismus.  Bio.  Z.  55,  45, 1913;  56,  174,  1913;  64,  106, 
1914. 

403.  SCHAUMANN,  H. :  Arch,  f .  Schiff.  u.  Tropenhyg.  Beih.  6.  July,  1914. 

404.  KARR,  W.  G. :  The  influence  of  water-soluble  vitamine  on  the  nutrition 

of  dogs.  Proc.  Soc.  Exp.  Biol.  Med.  17,  84,  1920;  J.  Biol  Chem. 
44,  255,  1920. 

405.  STILLING  AND  v.  MERING:  Ueber  experimentelles  Erzeugen  der  Osteo- 

malacie.     Z.  f.  d.  med.  Wiss.  803,  1889. 

406.  HEBRANT,  G.  AND  ANTOINE,  G. :  Sur  Tosteomalacie   du  chien.     Acad. 

de  meU  Beige.  28,  213,  1914. 

407.  GUERIN:  Cited  according  to  Cautley.  Rickets.    Garrod,  Batten  und 

Thursfield.     Diseases  of  Children,     p.  107.  London,  1913. 
407a.  ROLOFF,    F. :   Ueber   Osteomalazie   und   Rachitis.    Virch.   Arch.   37, 

1866;  Arch.  f.  wiss.  Thierh.  1,  189,  1875;  5,  152,  1879. 
407b.  REIMERS  AND  BOYE:  Ein  Beitrag  zur  Lehre  von  der  Rachitis.     Z.  f. 

innere  Med.  26,  953,  1905. 

408.  BULL,  :    J.  Comp.  Path.  31,  1918. 

408a.  MELLANBY,  E.  AND  MELLANBY,  M.:  The  experimental  production  ot 
thyroid  hyperplasia  in  dogs.  Proc.  Physiol.  Soc.;  J.  Physiol.  55, 
VII,  1921. 

408b.  MELLANBY,  E.  AND  MELLANBY,  M.:  The  application  of  results  ob- 
tained in  experiments  on  the  hyperplasia  of  dogs'  thyroids  to  the 
treatment  of  exophthalmic  goiter  (Graves' Disease).  Ibid.  55,  X, 
1921. 

408c.  HENDERSON,  P.  S. :  On  the  creatine  content  of  muscles  in  rickets  in 
dogs.  Proc.  Physiol.  Soc.  Oct.  19,  1918  ;  J.  Physiol.  Vol.  82. 


420  THE    VITAMINES 

408d.  FINDLAY,  L.,  PATON,  D.  N.  AND  SHARPE,  J.  S. :  Metabolism  of  rickets. 
Quart.  J.  Med.  14,  352,  1921. 

409.  HENRI QUES  AND  ANDERSEN:  Ueber  parenterale  Ernahrung  durch  intra- 

venose  Injektionen.     H.  88,  357,  1913. 

410.  FINGERLING,  G. :  Landw.  Vers.  Stat.  86,  75,  1915. 

411.  HOARE,  E.  W. :  System  of  veterinary  medicine.     Chicago  2,  1290,  1915. 

412.  JONES,  F.  S.  AND  ARNOLD,  J.  F. :  Staggers  in  sheep   in  Patagonia.     J. 

Exp.  Med.  26,  No.  6,  Dec.,  1917. 

413.  REID  AND  ASTON:  J.  New  Zealand.     Dept.  Agr.  15th  Nov.,  1910. 

414.  IBELE,   J. :  Die  Kontrolle  von   "Lecksucht"   und  falscher  Lecksucht 

beim  Renntier.  Mitt,  der  Vereins  z.  Ford.  d.  Moorkultur  im 
Deutsch.  Reiche.  36,  14,  1918. 

415.  EVVARD,  J.  M. :  Proc.  Iowa  Acad.  Sci.  22,  375,  1915. 

416.  HART,  E.  B.,  McCoLLUM,  E.  V.  AND  FULLER,  J.  G. :  The  role  of  inorganic 

phosphorus  in  the  nutrition  of  animals.  Amer.  J.  Physiol.  33, 
246,  1908. 

417.  HART  AND  MCCOLLUM:  The  influence  of  restricted  rations  on  growth. 

Proc.  Amer.  Soc.  Biol.  Chem.  J.  Biol.  Chem.  17,  II,  1914. 

418.  MCCOLLUM:  The  value  of  the  proteins  of  the  cereal  grains  and  of  milk 

for  growth  of  the  pig,  and  the  influence  of  the  plane  of  intake  on 
growth.  J.  Biol.  Chem.  19,  323,  1914. 

419.  HART  AND  MCCOLLUM:  Influence  on  growth  of  rations  restricted  to  the 

corn  or  wheat  grain.     Ibid.  19,  373,  1914. 

420.  HART  AND  STEENBOCK,  H. :  Maintenance  and  production  value  of  some 

protein  mixtures.     Ibid.  38,  267,  1919. 

421.  The   same.     Maintenance   and   reproduction   with    grains    and    grain 

products  as  the  sole  dietary.     Ibid.  39,  209,  1919. 

422.  KLEIN:  Milchw.  Zentralh.  43,  452,  1914;  44,  81,  1915;  44,  97,  1915. 

422a.  PLIMMER,  R.  H.  ADERS:  Note  on  "scurvy"  in  pigs.  Biochem.  J.  14,  570, 
1920. 

423.  GREEN,  H.  H. :  Dietetic  deficiency.     S.  Afr.  J.  Sci.  12,  289,  1916. 

424.  HART,  MILLER,  W.  S.  AND  MCCOLLUM:  Further  studies  on  the  nutritive 

deficiencies  of  wheat  and  grain  mixtures  and  the  pathological 
conditions  produced  in  swine  by  their  use.  J.  Biol.  Chem.  25,  239, 
1916. 

425.  ROBERTS,  G.  A. :  Effect  of  feeding  cottonseed  and  its  products  to  swine. 

J.  Amer.  Vet.  Med.  Ass.  49,  12,  1916. 

426.  ROMMEL,  G.  M.  AND  VEDDER,  E.  B. :  Beriberi  and  cottonseed  poisoning. 

in  pigs.     Prelim.  Comm.  J.  Agr.  Res.  5,  489,  1915. 

426a.  ZILVA,  S.  S.,  GOLDING,  J.,  DRUMMOND,  J.  C.  AND  COWARD,  K.  H.: 
Relation  of  the  fat-soluble  factor  to  rickets  and  growth  in  pigs. 
Biochem.  J.  15,427,  1921. 

427.  FRIEDBERGER,  FRANZ  AND  FROEHNER,  EUGEN:  Veter.  Path.  6th  Ed. 

Ed.  Chicago  2,  1908. 

428.  HUTYRA,  FRANCIS  AND  MAREK,  JOSEF:  Pathology  and  Therapeutics  of 

the  Diseases  of  Domestic  Animals.     Chicago  1,  1916. 


LITERATURE    TO    THE    TEXT  421 

429.  SCHEUNERT,     A.,     SCHATTKE,     A.     AND     L6TSCH,     E.  I      CaO,      MgO,      P2O6 

gehalt  von  Heu  und  Hafer,  nach  deren  Verfiitterung   Pferde   an 
Osteomalacie  erkrankten.     Bio.  Z.  36,  240,  1911. 

430.  KAWAKAMI,  Z. :  Disease  similar  to  beriberi,  produced  in  the  horse  by 

food-foundering.     Kyoto  Igaku  Zasshi.  14,  1,  1917. 

431.  STEWART:  A  report  on  the  disease  affecting  cattle  in  the  Moruya  District. 

New  S.  Wales  Agr.  J.,  1899. 

432.  LOTSCH,   E. :  Ueber  den  Stallmangel,  eine  eigenartige  Rindererkran- 

kung  in  sachsischen  Erzgebirge.     Zut  Kenntnis  des  Mineralstoff- 
wechsels.  Z.  f.  Inf.  u.  Hyg.  d.  Haustiere.  12,  205,  1912. 

433.  ASTON,  B.  C.  AND  REAKES,  C.  J. :  J.  New  Zealand  Dep.  Agr.  Nov.,  1911; 

Aug.,  1912;  Apr.,  1913;  Febr.,  1914. 

434.  RUSSELL,  H.  L.  AND  MORRISON,  F.  B. :  Univers.  Wise.  Agr.  Exp.  Sta. 

Bull.,  302,  55,  1919. 

435.  FLEISCHMANN,  FRITZ  :  Veranderungen  welche  bei  der  Dtirrheubereitung 

im  Grase  vor  sich  gehen.     Landw.  Vers.  Stat.  76,  237,  1912. 

436.  HENRY,    MAX:    Mortality  of    cattle  in    the    Bega    District    of    New 

South  Wales.     Veter.  J.  71,  No.  476,  Febr.,  1915. 

437.  FORBES,  E.  B. :  Mineral  feeds  for  farm  animals.    Annual  Meet.  Amer. 

Feed  Manuf.  Ass.  Chicago,  June  17-8,  1920. 

438.  PLACE  :  Diet  deficiency  and  disease  in  live  stock.     The  Farmers  Weekly 

5,  No.  125,  July,  1913. 

439.  THEILER,  A. :  Facts  and  theories  about  Stijfziekte  and  Lamziekte  Agr. 

J.  Union  of  S.  Africa,  1912. 

440.  HEDINGER,  E. :  Pathological  investigation  into  Lamziekte.     Dep.  Agr. 

Un.  S.  Africa,  1915. 

441.  VILJOEN,  P.  R. :  Investigation  into  Lamziekte  in  cattle.    V  and  VI. 

Dir.  Rep.  Veter.  Res.  Un.  S.  Africa  257,  1918. 

442.  SPRUELL:  Lamziekte  on  the  Cap  Plateau.    Agr.  J.  May,  1908. 

443.  MITCHELL,  D.  F.:  "Lamziekte"  II  Rep.    Dir.  Veter.  Res.  Dep.  Agr. 

Un.  S.  Africa,  1912. 

444.  WALKER,  JAMES:  Investigations  into  the  disease  Lamziekte  in  cattle. 

Ibid. 

445.  FUNK,  CASIMIR:  Results  of  studies  in  vitamines  and  deficiency  diseases 

during  the  years  1913-5.    Biochem.  Bull.  4,  304,  1915. 

445a.  HART,  E.  B.,  STEENBOCK,  H.  AND  HOPPERT,  C.  A.:  The  comparative 
influence  of  green  and  dried  plant  tissue,  cabbage,  orange  juice  and 
cod  liver  oil  on  calcium  metabolism.  Science  52,  318,  1920;  J. 
Biol.Chem.48,33, 1921. 

446.  STEAD:  Some  clinical  reflections  concerning  Lamziekte.     Agr.  J.  Un. 

S.  Africa  5,  386,  1913;  Ibid.  725,  1914;  July,  9,  1914. 

447.  THEILER,  A.,  GREEN,  H.  H.,  DU  TOIT,  P.  J.,  MEIER,  H.  AND  VILJOEN,  P. 

R. :  The  cause  and  prevention  of  Lamziekte.    Agr.  J.  Un.  S.  Africa 
Reprint  13,  July,  1920. 

448.  SHIGA,  K.  AND  KUSAMA,  SH.  :  Arch,  f .  Schiffs-  u.  Tropenhyg.     15,  Beih. 

3,  1911. 

449.  NOE:  Bull  soc..  path,  exotique.  p.  315,  1911. 


422  THE    VITAMINE6 

450.  MCCARRISON:    Pathogenesis    of    deficiency    diseases.     VII.  Effects    of 

autoclaved  rice  dietaries  on  gastro-intestinal  tract  of  monkeys. 
Ind.  J.  Med.  Res.  7,  283,  1919;  VIII.  7,  308,  1919;  Br.  Med.  J.  I. 
249,  Febr.  21st,  1920. 

451.  Hart,  Carl:  Ueber  die  experimentelle  Erzeugung  der  Moller-Barlowscher 

Krankheit  und  ihre  endgiiltige  Identifizierung  mit  dem  klassischen 
Skorbut.  Virch.  Arch.  208,  367,  1912. 

452.  HART,  C.  AND  LESSING,  OSCAR  :   Der  Skorbut  der  kleinen  Kinder.    Ferd. 

Enke.  Stuttgart  1913. 

453.  TALBOT,  TODD  AND  PETERSON:  Boston  Med.  &  Surg.  J.  169,  232,  1913. 

454.  HARDEN  AND  ZILVA:  Experimental  scurvy  in  monkeys.     J.  Path.  Bact. 

22,  246,  1919. 

455.  HARDEN  AND  ZILVA:  The  antiscorbutic  requirement  of   the    monkey. 

Biochem.  J.  14,  131,  1920. 

456.  ZILVA,  S.  S.  AND  STILL,  G.  F. :    Orbital  hemorrhage  with  proptosis  in 

experimental  scurvy.     Lane.  I.  May  8th,  1008,  1920. 

457.  HOWARD,  C.  P.  AND  INGVALDSEN,  F. :  The  mineral  metabolism  of  experi- 

perimental  scurvy  of  the  monkey.  Bull.  Johns  Hopk.  Hosp.  28, 
222,  1917. 

458.  HARDEN  AND  ZILVA:  Oedema  observed  in  a  monkey.     Lane.  II,  197, 

780,  1919. 

459..  HEWLETT,  TANNER  R.  AND  DE  KORTE:   Oedema   in  a  monkey.     Br. 
Med.  J.  II,  201,  1907. 

460.  CHICK,  HARRIETTS  AND  HUME,  ELEANOR  MARGARET:  The  production 

in  monkeys  of  symptoms  closely  resembling  those  of  pellagra  by 
prolonged  feeding  on  a  diet  of  low  protein  content.  Biochem.  J.  14, 
135,  1920. 

461.  HESS,  ALFRED  F. :  Infantile  scurvy.     Its  influence  on  growth  (length 

and  weight).    Amer.  J.  Dis.  Childr.  12,  152,  1916. 

462.  CHICK,  HARRIETTS  AND  DALYELL,  ELSIE    J. :  88th  Annual   Meeting. 

Br.  Med.  A.  Cambridge,  June  30- July  2nd,  1920,  Br.  Med.  J.  July 
31st,  147,  1920. 

462a.  SEIDELL,  ATHERTON:    Chemistry  of  vitamines.  J.  Ind.  Eng.  Chem. 
13,72,1921. 

463.  FUNK,  CASIMIR:  On  the  chemical  nature  of  the  substance  which  cures 

polyneuritis  in  birds  induced  by  a  diet  of  polished  rice.  J. 
Physiol.  43,  395,  1911. 

464.  SCHAUMANN,   H.  I  Zu    dem    Problem    der    Beriberiatiologie.     Arch.   f. 

Schiffs-  u.  Tropenhyg.  16,  825,  1912. 

465.  FUNK,  CASIMIR:  Studies  on  beriberi.    VII.  Further  facts  on  the  chem- 

istry of  the  vitamine-fraction  from  yeast  and  rice-polishings.  J. 
Physiol.  46,  173,  1913. 

466.  SUZUKI,  SHIMAMURA  AND  ODAKE:  Ueber  Oryzanin,  ein  Bestandteil  der 

Reiskleie  und  seine  physiologische  Bedetitung.     Bio.  Z.  43,  89,  1912. 

467.  SUZUKI  AND  MATSUNAGA:  On  the  occurrence  of  nicotinic  acid  in  rice 

bran.     Biochem.  Bull.  2,  228,  1913. 


LITERATURE   TO    THE    TEXT  423" 

468.  SCHAUMANN,  H. :  Ueber  die  Darstellung  und  Wirkungsweise  einer  der 

in  der  Reiskleie  enthaltenen  gegen  experimentelle  Polyneuritis 
wirksamen  Substanz  (Vorl.  Mitt).  Arch.  f.  Schiffs-  u.  Tropenhyg. 
16,  349,  1912;  D.  med.  W.  No.  26,  1255,  1912. 

469.  WELLMAN,  C.,  EUSTIS,  A.  C.  AND  SCOTT,  L.  C. :  Rapid  cure  of  polyneu- 

ritis  gallinarum  by  intramuscular  injection  of  a  substance  isolated 
from  rice.  Amer.  J.  Trop.  Dis.  &  Prev.  Med.  1,  295,  1913. 

470.  EUSTIS,  A.  C.  AND  SCOTT,  L.  C. :  Isolation  of  vitamine  from  rice-polish- 

ings.     Biochem.  Bull.  3,  466,  1914. 

471.  VEDDER,  E.  B.  AND  WILLIAMS,  R.   R. :  Beriberi-preventing  substances 

or  vitamines  contained  in  rice  polishings.  VI.  Philipp.  J.  Sci.  8 
(B),  175,  1913. 

472.  WILLIAMS,  R.  R.   AND  SALEEBY,   N.  B. :   Experimental    treatment  of 

human  beriberi  with  constituents  of  rice  polishings.  Ibid.  10(B), 
99,  1915. 

473.  WILLIAMS,  R.  R. :  Chemistry  of  the  vitamines.     Ibid.  11  (A),  49,  1916. 

474.  ISSOGLIO,  GIOVANNI:  Chemical  composition  of  the  by-products  of  the 

working  up  of  the  rice.     Atti  acad.  sci.  Torino,  54,  980,  1919. 

475.  KONDO,  H.  AND  GOMI,  N. :   Efficacious  constituents   of  rice  bran.     J. 

Pharm.  Soc.  Japan  No.  391,  1013,  1913. 

476.  MURAI,  T. :  A  new  method  for  the  extraction  of  the  efficacious  constit- 

uents of  the  rice  bran.     Ibid.  No.  386,  1914. 

477.  TSUZUKI,  J. :  Remedy  for  beri-beri.    Brit.  25,322,  Nov.  5th,  1912. 

478.  GAMS,  A.  AND  SCHREIBER,  B.:  Vitamines.    U.  S.  P.  1,235,198,  July  31, 

1917;  Soc.  Anon,  pour  1'Ind.  chim.  a  Bale.  Schweiz.  76,  494,  Apr. 
1st,  1918. 

479.  BRILL,  HARVEY  C. :  The  antineuritic  properties  of  the  infusorial  earth 

extract  of  the  hydrolyzed  extract  of  rice  polishings.  Philipp.  J. 
Sci.  12,  199,  1917. 

480.  FRASER,  H.  AND  STANTON,  A.  T. :  The  chemistry   of    rice-polishings. 

Lane.  I,  1021,  1915. 

481.  DRUMMOND,  J.  C.  AND  FUNK,  CASIMIR:  Chemical  investigation  of  the 

phosphotungstic  acid  precipitate  fron  rice  polishings.  Biochem. 
J.  8,  598,  1914. 

482.  BARGER:  Simpler  Natural  Bases.    Longmans,   Green  &  Co.,  London, 

1914. 

483.  HOFMEISTER,  FRANZ  AND  TANAKA,  M. :  cited  by    Hofmeister,  Erg.  d. 

Physiol.  16,  510,  1918. 

484.  HOFMEISTER,  FR.  :  Zur  Kenntnis  der  alkaloidischen  Bestandteile  der 

Reiskleie.    Bio.  Z.  103,  218,  1920. 

485.  FUNK,  CASIMIR  :  Further  facts  concerning  the  chemistry  of  the  vitamine- 

fraction  from  yeast.     Brit.  Med.  J.  Apr.  19th,  1913. 

486.  FOLIN,  O.  AND  MACALLUM,  A.  B.,  JR.:  J.  Biol.  Chem.  11,  265,  1912;  13, 

363,  1912. 

487.  BARSICKOW:  Experimentelle  Untersuchungen  iiber  die  therapeutische 

Wirkung  der  Hefe  bei  der  alimentaren,  multiplen  Polyneuritis  der 
Meerschweinchen  und  Tauben.  Bio.  Z.  48,  418,  1913. 


424  THE    VITAMINES 

488.  EDIE,  EVANS,  MOORE,  SIMPSON  AND  WEBSTER:  The  antineuritic  bases 

of  vegetable  origin  in  relationship  to  beriberi,  with  a  method  of 
isolation  of  torulin,  the  antineuritic  base  of  yeast.  Biochem.  J.  6, 
234,  1913. 

489.  FUNK,    CASIMIR:   Fractionation    of   phosphotungstic    acid   precipitate 

with  acetone  as  a  useful  method  for  the  preparation  of  the  vitamine- 
fraction  from  yeast.  Biochem.  Bull.  5,  1,  1916;  U.  S.  P.  1,162,908, 
Dec.  7th,  1915. 

490.  VAN  SLYKE,  D.D  .:  J.  Biol.  Chem.,  30,  115,  1917. 

491.  BOHRINGER,  C.  F.  AND  SONS:  Separating  active  substances  from  food 

and  organ  extracts.     Holl.  2600,  Oct.  15,  1918. 

492.  COOPER,  E.  A.:  The  curative  action  of  autolyzed  yeast  against  avian 

polyneuritis.     Biochem.  J.  8,  250,  1914. 

493.  FUNK,  CASIMIR  AND  DUBIN,  HARRY  E. :  A  test  for  antiberiberi  vitamine 

and  its  practical  application.     J.  Biol  Chem.  44,  487,  1920. 

494.  DRUMMOND,   J.   C. :  Observations  on  the  phosphotungstates  of  certain 

bases  and  aminoacids.     Biochem.  J.  12,  5,  1918. 

495.  SEIDELL,   ATHERTON:   Vitamines   and   nutritional   diseases.     A   stable 

form  of  vitamine,  efficient  in  the  prevention  and  cure  of  certain  nu- 
tritional deficiency  diseases.  Repr.  325,  U.  S.  P.  H.  S.  Rep.  Febr. 
18th,  1916;  Seidell  A.,  Vitamines,  U.  S.  Pat.  1,173,317,  Febr.  28th, 
1916. 

496.  WILLIAMS,  R.  R.  AND  SEIDELL,  A.:  The  chemical  nature  of  the  "vita- 

mines".      II.  Isomerism   in   natural   antineuritic   substances.     J. 

Biol.  Chem.  26,  431,  1916. 
496a.  OSBORNE,  T.  B.  AND  LEAVENWORTH,  C.  L.:    Effect  of  alkali  on  the 

efficiency  of  water-soluble  vitamine  B.    J.  Biol.  Chem.  45,  423,  1921. 
496b.  RHODEHAMEL,  H.  W.  AND  STUART,  A.  H.:    Atropine  sulphate  from 

Datura  stramonium.     J.  Ind.  Eng.  Chem.  13,  48,  1921 . 
496c.  SEIDELL,  A. :  A  stable  silver  vitamine  compound  obtained  from  brewer's 

yeast.    Publ.  Health  Rep.  36,  665,  1921. 

497.  VOEGTLIN,  C.  AND  WHITE,  G.  F. :   Can  adenine  acquire  antineuritic 

properties?  J.  Pharmacol.  9,  155,    1916. 

498.  DRUMMOND,  J.  C. :  A  study  of  water-soluble  accessory  growth  promoting 

substance  in  yeast.     I.  Biochem.  J.  11,  255,  1917. 

499.  ABDERHALDEN,  E.  AND  SCHAUMANN,  H. :   Beitrag   zur   Kenntnis  von 

organischen  Nahrungsstoffen  mit  spezifischer  Wirkung.  Arch.  f. 
Physiol.  (Pfl.  Arch.)  172,  1,  1918. 

500.  ABDERHALDEN,   E. :  Weitere  Beitrage  zur  Kenntnis  von   organischen 

Nahrungsstoffen  mit  spezifischer  Wirkung.     Ibid.  178,  260,  1920. 

501.  SUGIURA,   KANEMATSU:  A  preliminary  report  on  the  preparation  of 

antipolyneuritic  substances  from  carrots  and  yeast.  J.  Biol.  Chem. 
36,  191,  1918. 

502.  OSBORNE,  T.  B.  AND  WAKEMAN,  A.  J. :  Extraction  and  concentration  of 

the  water-soluble  vitamine  from  brewer's  yeast.  Ibid.  40,  383, 
1919. 

503.  MYERS,  C.  N.  AND  VOEGTLIN,  C. :    The  chemical  isolation  of  vitamine. 

Ibid.  42.  199,  1920. 


LITERATURE   TO   THE   TEXT  425 

504.  POL,  D.  J.  HULSHOFF:  X-acid  as  a  remedy  in  polyneuritis  and  beriberi. 

J.  Physiol.  51,  432,  1917;  Ned.  Tijdsr.  v.  Gen.  11,  806,  1917. 

505.  VOEGTLIN,  C.  AND  TowLEs:   The  treatment  of  experimental  beriberi 

with  extracts  of  spinal  cord.     J.  Pharmacol.  5,  67,  1913. 

506.  SULLIVAN,  M.X.  AND  VOEGTLIN,  C.:  Distribution  in  foods  of  the  so-called 

vitamines  and  their  isolation.     Proc.  Soc.  Biol  Chem.,  Dec.  27th, 
1915;  J.  Biol.  Chem.  24,  XVI,  1956. 

507.  STEENBOCK,  H. :  Antineuritic  substance  from  egg  yolk.     Proc.  Amer. 

Soc.  Biol.  Chem.  J.  Biol.  Chem.,  29,  XXVII,  1917;  J.  Biol.  Chem. 
29,  495,  1917. 

508.  WILLIAMS,  R.  R. :  The  chemical  nature  of  the  "vitamines."    I.  Anti- 

neuritic properties  of  the  hydroxypyrimidines.     J.  Biol.  Chem.  25, 
437,  1916. 

509.  The  same.     Structure  of  antineuritic  hydroxy-pyridines.     Proc.  Soc. 

Exp.  Biol.  Med.  14,  25,  1916. 

510.  HARDEN,    A.  AND  ZILVA,  S.  S. :    Alleged    antineuritic    properties    of 

o-hydroxypyridines  and  adenine.     Biochem.  J.  11,  172,  1917. 

511.  DUTCHEB,  R.  ADAMS,  HOLM,  C.  E.  AND  BIERMAN,  HARLOW:    Further 

observations  on  the  antineuritic  properties  of  chemical  substances. 
Science  52,  589,  1920. 

512.  FUNK,  CASIMIR:  The  influence  of  radium  emanation  on  the  activity  of 

vitamines.    Proc.  Soc.  Exp.  Biol.  Med.  14,  9,  1916. 

513.  ZILVA,  SOLOMON  SYLVESTER:  The  action  of  ultra-violet  rays  on  the 

accessory  food  factors.    Biochem.  J.  13,  164,  1919. 

514.  SUGIURA,  KANEMATSU  AND  BENEDICT,  STANLEY  R. :  The  action  of  radium 

emanation  on  the  vitamines  of  yeast.     J.  Biol  Chem.  39,  421,  1919. 

515.  WEILL,  E.  AND  MOURIQUAND,  G. :  Action  des  rayons  X  sur  grains  d'orge 

et  carence.    Soc.  biol.  9  Oct.,  1918. 

516.  VOEGTLIN,  C.  AND  MYERS,  C.  N. :   Phosphorus  as  an  indicator  of  the 

vitamine  content  of  corn  and  wheat  products.     U.  S.  Publ.  Health 
Serv.  Repr.  No.  471,  49,  1918. 

517.  GREEN,  HENRY  H. :  The  vitamine  content  of  maize  and  maize-milling 

products,  and  the  ambiguity  of  its  correlation  with  the  phosphoric 
acid  content.     S.  Afr.  J.  Sci.  14,  519,  1918. 

518.  OTTOW,  W.  M. :  Examination,  preservation  and  treatment  of  the  silver 

skin  of  rice.     Gen.  Tijdsr.  v.  Ned.  Indie,  55,  75,  1915. 

519.  FUNK,  CASIMIR:  An  attempt  to  estimate  the  vitamine-fraction  in  milk. 

Biochem.  J.  7,  211,  1913. 

520.  BRILL,  HARVEY  C.  AND  ALINCASTRE,  CECILIO:   The  possible  maximum 

vitamine  content  of  some  Philippine  vegetables.     Philipp.  J.  -Sci. 
12  A,  127,  1917. 

521.  SEIDELL,  A. :  The  vitamine  content  of'brewers,  yeast.     J.  Biol.  Chem. 

29,  145,  1917. 

522.  EDDY,   W.   H. :  Further  observations  on  pancreatic  vitamine.     Proc. 

Soc.  Exp.  Biol.  Med.  14,  164,  1917. 


426  THE    VITAMINES 

523.  FUNK,  CASIMIR  AND  MACALLUM,  A.  B.,  JR.:   On  the  chemical  nature  of 

substances  from  alcoholic  extract  of  various  foodstuffs  which  give 
a  color  reaction  with  phosphotungstic  and  phosphomolybdic  acids. 
Biochem.  J.  7,  356,  1913. 

524.  LEWIS,  HOWARD  B.  AND  NICOLET,  BEN  H. :  The  reaction  of  some  purine, 

pyrimidine  and  hydantoin  derivatives  with  the  uric  acid  and  phenol 
reagents  of  Folin  and  Denis.  J.  Biol.  Chem.  16,  369,  1913. 

525.  FUNK,  CASIMIR:  The  influence  of  the  milling  of  maize  on  the  chemical 

composition  and  the  nutritive  value  of  maize-meal.  J.  Physiol. 
47,  389,  1913. 

526.  GREEN,  HENRY,  H. :  Experimental  expression  of  the  relationship  between 

the  content  of  foodstuff  in  antineuritic  hormone  and  the  period  of 
healthy  survival  of  animals  upon  it.  S.  Afr.  J.  Sci.  14,  483,  1918. 

527.  EDDY  AND  STEVENSON,  H.  C. :    Further  studies  in  the  measurement  of 

the  vitamine  content.  Proc.  Soc.  Exp.  Biol.  Med.  17,  52,  122,  1920; 
J.  Biol.  Chem.  43,  295,  1920. 

528.  WILLIAMS,  R.  J. :  A  quantitative  method  for  determination  of  vitamine. 

J.  Biol  Chem.  42,  259,  1920. 

529.  DE  SOUZA,  GERALDO  PAULA  AND  McCoLLUM,  E.  V. :    A  study   of  the 

factors  which  interfere  with  the  use  of  yeast  as  test  organism  for 
the  antineuritic  substance.  Ibid.  44,  113,  1920. 

530.  FUNK,  CASIMIR  AND  MACALLUM,  A.  B.,  JR.  :  The  action  of  yeast  fractions 

on  the  growth  of  rats.     J.  Biol.  Chem.  27,  63,  1916. 

531.  EMMETT,  A.  D.  AND  STOCKHOLM,  MABEL:   Water-soluble  vitamine.     II. 

The  relation  of  the  antineuritic  and  water-soluble  B-vitamines  to 
the  yeast  growth-promoting  stimulus.  Ibid.  43,  287,  1920. 

531a.  GOY,  PIERRE.:  Les  v£getaux  inferieurs  et  les  facteurs  assessoires  de 
la  croissance.  C.  r.  172,  242,  1921. 

531b.  FRANKEL,  S.  AND  SCHWARTZ,  E. :    Biochem.  Z.  112,  203,  1920. 

532.  EIJKMAN,   C. :  Ueber  die    Natur   und   Wirkungsweise    der  gegen    die 

experimentelle  Beriberi  wirksamen  Substanzen.  Arch,  f .  Schiff-  u. 
Tropenhyg  17,  328,  1913. 

533.  FUNK,   CASIMIR:   Fortschritte   der  experimentellen  Beriberiforschung 

in  den  Jahren  1911-1913.     M.  med.  W.  No.  36,  1913. 

534.  FUNK:  Further  experimental  studies  on  beri-beri.     The  action  of  certain 

purine-  and  pyrymidine-derivatives.     J.  Physiol.  45,  489,  1913. 

535.  COOPER,  E.  A.:  The  preparation  from  animal  tissues  of  a  substance 

which  cures  polyneuritis  in  birds  induced  by  diets  of  polished  rice. 
Biochem.  J.  268,  1913. 

536.  COOPER:  On  the  protective  and  curative  properties  of  certain  foodstuffs 

against  polyneuritis  induced  in  birds  by  a  diet  of  polished  rice. 
Part  I,  J.  of  Hyg.  12,  436,  1913. 

537.  ABDERHALDEN,  E.  AND  EWALD,  G. :   Gibt  es  lebenswichtige  unbekannte 

Nahrungstoffe?    Z.  f.  ges.  exp.  Med.  5,  1,  1916. 

537a.  SEAMAN,  EMILY  C. :  Influence  of  alcoholic  extract  of  thyroid  gland 
upon  polyneuritic  pigeons  and  the  metamorphosis  of  tadpoles. 
Amer.  J.  Physiol.  53,  101,  1920. 


LITERATURE    TO   THE   TEXT  427 

538.  RAMOINO,  P.:  Incomplete  diets.    Researches  on  gaseous  exchange  on 

subjects  receiving  a  rice  diet.    Arch.  ital.  biol.  65,  1,  1916. 

539.  JANSEN,  B.  C.  P.,  AND  MANGKOEWINOTO,  R.  M.  M. :  Medel  v.  d.  Burg. 

Gen.  Dienst.  Ned.  Indie,  No.  31,  51,  1920. 

540.  MASSALONGO,  R. :  Acute  polyneuritis,  a  form  of  beriberi.     Contribution 

to  a  theory  of  vegetable  hormones.     Riv.  med.  32,  1316,  1916. 

541.  LUMIERE,  A.:  Bull,  de  PAcad.  de  MeU  83,  310,  1920;  Presse  me*d.  28,  29, 

1920. 

541a.  DAMIANOVICH,  H.  L.  AND  PILADO,  MATHAU  C.:  Biochemical  and 
clinical  research  on  vitamines.  Rev.  de  la  Ass.  med.  Arg.  34,  279, 
286,303,  1921. 

542.  DUTCHER,  R.  ADAMS  :  Vitamine  studies  I.    Observations  on  the  catalase 

activity  of  tissues  in  avian  polyneuritis.  J.  Biol.  Chem.  36,  63, 
1918. 

543.  DUTCHER  AND  COLLATZ:  II.  Ibid.  36,  547,  1918. 

544.  BURGE,  W.  E.  AND  NEILL,  A.  J. :    Effect  of  starvation  on  the  catalase 

content  of  the  tissues.    Amer.  J.  Physiol.  43,  58,  1917. 

545.  JANSEN,  B.  C.  P.:  Is  vitamine  identical  with  secretin?    Gen.  Tijdsr. 

v.  Ned.  Indie.  58,  191,  1918. 

546.  VOEGTLIN,  C.  AND  MYERS,  C.  N. :    Comparison  of  influence  of  secretin 

and  antineuritic  vitamine  on  pancreatic  secretion  and  bile  flow. 
J.  Pharmacol.  13,  301,  1919. 

546a.  COWGILL,  G.  R.:  Studies  in  the  physiology  of  vitamines.  Is  water- 
soluble  vitamine  identical  with  secretin?  .Proc.  Soc.  Exp.  Biol. 
Med.  18, 148, 1921.  Does  vitamine  B  stimulate  glands  in  a  manner 
similar  to  the  alkaloid  pilocarpine.  Ibid.  18,  290, 1921. 

546b.  ANREP,  G.  V.  AND  DRUMMOND,  J.  C.:  Note  on  supposed  identity  of 
vitamine  B  and  secretin.  J.  Physiol.  54,  349,  1921. 

547.  BICKEL,  A.:  Bin  neues  Pflanzensekretin.    Berl.   klin.  W.  54,  74,  552, 

1917.    Eisenhardt,  W.,  54,  553,  1917.    Djenab,  K,  54,  624,  1917. 

548.  VAN  EWEYK  :  Die  Wirkung  des  Spinatextraktes  auf  die  innere  Pankreas- 

sekretion.    Virch.  Arch.  227,  113,  1920. 

549.  BORUTTAU,  H.:  Das  Verhalten  der  Erganzungsstoffe.     II.   Spezifische 

antidiabetische  Substanzen.    Bio.  Z.  88,  420,  1918. 

550.  UHLMANN,  F. :  Beitrag  zu  der  Pharmakologie  der  Vitamine.     Z.  f .  Biol. 

68,  419,  1918;  68,  457,  1918. 

551.  BRADDON,  L.,  AND  COOPER,  E.  A.:  J.  Hyg.  14,  331,  1914. 

552.  MUCKENFUSS,  A.  M. :  The  presence  of  food  accessories  in  urine,  bile,  and 

saliva.  J.  Amer.  Chem.  Soc.  40,  1606,  1918.  The  excretion  of 
vitamines.  Arch  of  Pediatr.  36,  80,  1919. 

553.  GAGLIO,  G. :  Vitamine  in  urine.     Policlinico.  26,  1381,  1919. 

554.  OSBORNE,  T.  B.  AND  MENDEL,  L.  B.  i  The  role  of  vitamines  in  the  diet. 

J.  Biol.  Chem.  31,  149,  1917. 

555.  MAURER:  Polyneuritis  der  Htihner  und  Beriberi,  eine  chronische  Oxal- 

saurevergiftung.    M.  med.  W.  54,  731,  1907. 


428  THE    VITAMINES 

556.  CHAMBERLAIN,  BLOOMBEEGH  AND  KILBOURNE  :  A  study  of  the  influence 

of  the  rice  diet  and  of  inanition  on  the  production  of  multiple 
neuritis  of  fowls  and  the  bearing  thereof  to  the  etiology  of  beri- 
beri. Philipp.  J.  Sci.  6  (B),  177,  1911. 

557.  EIJKMAN,  C.  AND  VAN  HooGENHUYZE,  C.  J.  C. :   Influence  of  feeding  and 

of  starvation  on  the  development  of  polyneuritis  gallinarum.  Proc. 
Akad.  Wetterschap.  18,  1467,  1916;  Virch.  Arch.  222,  301,  1916. 

558.  WALSHE,  F.  M.  R.:  The  deficiency  theory  of  the  origin  of  beriberi  in 

the  light  of  clinical  and  experimental  observations  on  the  disease, 
with  an  account  of  a  series  of  40  cases.  Quart.  J.  Med.  11,  320, 
1918. 

559.  FUNK,  CASIMIR:  Studies  on  beri-beri.    The  probable  role  of  vitamines 

in  the  process  of  digestion  and  utilization  of  food.  Proc.  Physiol. 
Soc.  Dec.  13th,  1913;  J.  Physiol.  Vol.  47. 

560.  FUNK:  Studien  iiber  Beriberi.    XI.  Die  Rolle  der  Vitamine  bein  Kohlen- 

.hydratstoffwechsel.     Ztschr.  f.  physiol.  Chem.  89,  378,  1914. 

561.  BRADDON,  L.  AND  COOPER,  E.  A.:  The  influence  of  the  total  fuel-value 

of  a  dietary  upon  the  quantity  of  vitamine  required  to  prevent 
beriberi.  Brit.  Med.  J.  June  20,  1914;  J.  Hyg.  14,  331,  1914. 

562.  WEILL,  E.  AND  MOURIQUAND,  G. :    Les  maladies  par  carence  (carence 

expe"rimentale,  carence  clinique).     Rev.  de  me'd.  No.  1  et  2,  1916. 

563.  VEDDER,  E.  B. :  Is  the  neuritis-preventing  vitamine  concerned  in  carbo- 

hydrate metabolism?    J.  Hyg.  17,  1,  1918. 

564.  MAIGNON,  F. :  Etudes  comparers  sur  Tinfluence  des  hydrates  de  carbone 

et  des  graisses  sur  le  pouvoir  nutritif  des  albumines  alimentaires. 
C.  r.  167,  172,  1919. 

565.  BIERRY,  H.  AND  PORTIER,  P. :  C.  r.  soc.  biol.  81,  574,  1918. 

566.  BIERRY,  H.:  C.  r.  169,  197,  1919. 

567.  JOHNS,  CARL  O.  AND  FINKS,  A.  J. :  Studies  in  nutrition  IV.    The  nutri- 

tive value  of  peanut  flour  as  a  supplement  to  wheat  flour.  J.  Biol. 
Chem.  42,  569,  1920. 

567a.  SCOTT,  E.  L.,  AND  HONEYWELL,  H.  E.:    Sugar  in  blood   of  normal 
pigeons.    Amer.  J.  Physiol.  54,  349, 1921. 

568.  FUNK,  CASIMIR:   Action  of  substances,  influencing  the  carbohydrate 

metabolism,  in  experimental  beriberi.     J.  Physiol.  53,  247,  1919. 

569.  ELIAS,  H.  AND  KOLB,  L. :  Die  Rolle  der  Saure  in  Kohlenhydratstoff- 

wechsel.     II.  Hungerdiabetes.  Bio.  Z.  52,  330. 

570.  OSBORNE  AND  MENDEL:  The  influence  of  cod  liver  oil  and  some  other 

fats  in  growth.     J.  Biol.  Chem.  17,  401,  1914. 

571.  Anon. :  Cod  liver  oil  industry.     Pharm.  Era  48,  255,  1915. 

572.  ISCOVESCO:  Lecithide  contenue  dans   Phuile  de  foie  de  morue.     C.  r. 

soc.  biol.  76,  I,  34;  II,  74,  II,  117,  1913;  10  et  17  Janv.  1914. 

573.  GAUTIER,  ARMAND  AND  MOURGUES:  C.  r.  107,  110  et  626,  1888. 

574.  FUNK,  CASIMIR  :  Biochemistry  of  cod-liver  oil.    Biochem.  Bull.  4, 365, 1915. 
574a.  ZILVA,  S.  S.  AND  MIURA,  M.:    A  note  on  the  activity  of  fat-soluble 

accessory  factors  in  cod  liver  oil  and  butter.    Lane.  I.  323,  1921. 

575.  PAAL,  C.,  AND  ROTH,  K. :  Reduktion  der  Fette.     B.  42,  1541,  1909. 


LITERATURE    TO    THE   TEXT  429 

576.  CHAPMAN,  A.  C. :  Spinacene.   A  new  hydrocarbon  from  certain  fish  oils. 

J.  Chem.  Soc.  Ill,  56,  1917. 

577.  TSUJIMOTO,  M. :  Squalene :  a  highly  unsaturated  hydrocarbon  in  shark 

liver  oil.     J.  Ind.  and  Eng.  Chem.  12,  63,  1919;  12,  73,  1919. 

578.  KUBOTA,  B. :  The  chemical  constitution  of  squalene.     Tokyo  Kwagaku 

Kwaishi.  39,  879,  1918. 

579.  BULL,  HENRIK:  The  composition  of  cod  liver  oil.    Tidskr.  Kemi  Farm. 

Terapi  14,  1916. 

580.  ROGERS,  L. :  Preparation  of  sodium  morrhuate.     Brit.  Med.  J.  Sept. 

27th,  1919. 

581.  OSBORNE  AND  MENDEL:  Further  observations  of  the  influence  of  natural 

fats  upon  the  growth.     J.  Biol.  Chem.  20,  379,  1915. 

582.  DRUMMOND,  J.  C. :  Researches  on  the  fat-soluble  accessory  substance. 

I.  Observations  upon  its  nature  and  properties.    Biochem.  J.  13, 
81,  1919. 

582a.  STEENBOCK,  H.,  SELL,  MARIANA  T.  AND  BUELL,  MARY  B.:  Fat- 
soluble  vitamine.  VII.  The  fat-soluble  vitamine  and  yellow 
pigmentation  in  animal  fats,  with  some  observations  on  its  stability 
tosaponification.  J.  Biol.  Chem.  47,  89, 1921. 

583.  McCoLLUM,  SIMMONDS  AND  PiTZ :  Distribution  of  the  fat-soluble  A,  the 

dietary  essential  of  butter  fat.     Amer.  J.  Physiol.  41,  361,  1916. 

584.  OSBORNE  AND  MENDEL:  The  extraction  of  " fat-soluble"  vitamine  from 

green  foods.     Proc.  Soc.  Exp.  Biol.  Med.  16,  98,  1919. 

585.  ZILVA,  S.  S. :  The  extraction  of  the  fat-soluble  factor  of  cabbage  and 

carrot  by  solvents.     Biochem.  J.  14,  494,  1920. 

586.  STEENBOCK,  H.  AND  BOUTWELL,  P.  W. :   Fat-soluble  vitamine  VI.    The 

extractability   of   the   fat-soluble  vitamine  from  carrots,  alfalfa, 
and  yellow  corn  by  fat  solvents.     J.  Biol.  Chem.  42,  131,  1920. 

587.  STEENBOCK,  H. :  White  corn  vs.  yellow  corn  and  a  probable  relation 

between   the   fat-soluble   vitamine    and   yellow   plant   pigments. 
Science  50,  352,  1919. 

588.  STEENBOCK  AND  BOUTWELL:  Fat-soluble  vitamine.    III.  The  compara- 

tive nutritional  value  of  white  and  yellow  maizes.     J.  Biol.  Chem. 
41,  81,  1920. 

589.  PALMER,  LEROY  S. :  Carotinoids  as  fat-soluble  vitamine.    Science,  50, 

501,  1919. 

589a.  PALMER,  LEROY  S.  AND  KENNEDY,  CORNELIA:  The  relation  of  plant 
carotinoids  to  growth  and  reproduction  of  albino  rats.  J.  Biol. 
Chem.  46,  559,  1921. 

590.  ROSENHEIM,   OTTO  AND  DRUMMOND,   J.   C.:    Relation  of  lipochrome 

pigments  to  fat-soluble  accessory  food  factors.    Lane.  I,  862,  1920. 

590a.  STEPHENSON,  M.:  Note  on  differentiation  of  yellow  pigments  from 
fat-soluble  vitamine.  Biochem.  J.  14,  715,  1920. 

590b.  STEENBOCK,  H.,  SELL,  M.  T.,  NELSON,  E.  M.  AND  BUELL,  M.  V. :  Fat- 
soluble  vitamine.  Proc.  Soc.  Biol.  Chem.  J.  Biol.  Chem.  46, 
XXXII,  1921. 

591.  VAN  DEN  BERGH,  A.  A.  HUMANS  AND  MULLER,  P. :   Serum-lipochrome. 

I.     Proc.  Acad.  Sci.  Amsterdam  22,  748,  1920. 


430  THE   VITAMINES 

592.  DRUMMOND,  J.  C.  AND  COWARD,  KATHARINE  HOPE:   Researches  on  the 

fat-soluble  accessory  substance.  V.  The  nutritive  value  of  animal 
and  vegetable  oils  and  fats  considered  in  relation  to  their  color. 
Biochem.  J.  14,  668,  1920. 

593.  McCoLLUM,  E.  V.,  SIMMONDS,  N.  AND  STEENBOCK,  H. :  A  method  for  the 

separation  of  the  dietary  essential  "fat-soluble  A",  from  butter 
fat.  Proc,  Amer.  Soc.  Biol.  Chem.;  J.  Biol.  Chem.  29,  XXVI,  1917. 

594.  STEENBOCK,  H.,  BOUTWELL,  P.  W.  AND  KENT,  HAZEL  E. :    Fat-soluble 

vitamine.  I.     J.  Biol.  Chem.  35,  517,  1918. 

595.  HOPKINS,  F.  G. :  Discussion  on  the  present  condition  of  vitamines  in 

clinical  medicine.    Brit.  Med.  J.  July  31st,  147,  1920., 

596.  MCCOLLUM,  E.  V.  AND  DAVIS,  M.:  Proc.  Soc.  Exp.  Biol.  Med.  11,  101, 

1913-1914. 

597.  DRUMMOND,  J.  C.:  The  fat-soluble  accessory  factor.     J.  Physiol.  52, 

344,  1919. 

598.  STEENBOCK  AND  BOUTWELL:  Fat-soluble  vitamine.  V.    Thermostability 

of  the  fat-soluble  vitamine  in  plant  materials.  J.  Biol.  Chem.  41, 
163,  1920. 

599.  OSBORNE  AND  MENDEL:  The  stability  of  the  growth-promoting  substance 

in  butter  fat.    Ibid.  24,  37,  1916. 
599a.  HOPKINS,  F.  G. :    Effects  of  heat  and  aeration  on  fat-soluble  vitamine. 

Biochem.  J.  14,  725,  1920. 
599b.  DRUMMOND,  J.  C.  AND  COWARD,  K.  H.:    Researches  on  fat-soluble 

accessory  factors  (vitamin  A).    VI.  Effect  of  heat  and  oxygen  on 

vitamin  value  of  butter.     Ibid.  14,  734,  1920. 

600.  The  same.    Nutritive  factors  in  plant  tissues.     IV.  Fat-soluble  vita- 

mines.     Ibid.  14,  549,  1920. 

600a.  ZILVA,  S.  S. :  Action  of  ozone  on  fat-soluble  factor  in  fats.  Biochem. 
J.  14,  740,  1920. 

601.  FAHRION,  W. :  Fett  und  Vitamine.     Chem.  Umschau  97,  109,  1920. 

602.  FREUDENBERG,  ERNST  AND  KLOCMAN,  LUDWIG:  Spasmophilia.  II.     J.  f. 

Kinderh.  79,  700. 

603.  HESS,  A.  F.  AND  UNGER,  LESTER  J. :  The  deleterious  effect  of  the  alkalin- 

ization  of  infants'  foods.      J.  A.  M.  A.  73,  1353,  1919. 

603a.  DRUMMOND,  J.  C.,  COWARD,  K.  H.  AND  WATSON,  A.  F.:  Notes  on  the 
factors  influencing  the  value  of  milk  and  butter  sources  of  vitamin 
A.  Biochem.  J.  15,  540, 1921 . 

604.  DRUMMOND,  J.  C.  AND  COWARD,  K.  H. :   Researches  on  the  fat-soluble 

accessory    substance.    III.     Technique    for    carrying    out    feeding 
tests  for  vitamine  A  (fat-soluble  A).    Biochem.  J.  14,  661,  1920. 
604a.  STAMMERS,  A.  D.:    The  value  of  steam  distilled  palm  kernel  oil  as  a 
control  fat.    Biochem.  J.  15,  489,  1921. 

605.  COWARD   AND   DRUMMOND:   Researches   on  the   fat-soluble    accessory 

substance.  IV.  Nuts  as  a  source  of  vitamin  A.  Biochem.  J.  14, 
665,  1920. 

605a.  DELF,  E.  MARION:  The  distribution  of  accessory  food  factors  (vita- 
mins) in  plants.  S.  Afr.  J.  Sci.  17, 121,  1920. 


LITERATURE   TO   THE   TEXT  431 

605b.  COWARD,  K.  H.  AND  DRUMMOND,  J.  C.:  Formation  of  vitamin  A  in 
living  plant  tissues.  Biochem.  J.  15,  530,  1921. 

605c.  HUGHES  J.  S.,:  Effect  of  fat-soluble  vitamine  content  of  the  feed  on 
the  fat-soluble  vitamine  content  of  adipose  tissue.  Science  52, 
565,  1920. 

605d.  DRUMMOND,  J.  C.,  GOLDING,  J.,  ZILVA,  S.  S.  AND  COWARD,  K.  H:  Nu- 
tritive value  of  lard.  Biochem.  J.  14,  742, 1920. 

606.  DRUMMOND,  J.  C. :  Researches  on  the  fat-soluble  accessory  substance. 

II.  Observations  on  its  role  in  nutrition  and  influence  on  fat  metab- 
olism. Ibid.  13,  95,  1919. 

607.  MENDEL,  LAFAYETTE  B. :  The  fat-soluble  vitamine.    N.  Y.  State  Med. 

J.  20,  212,  1920. 

608.  McCARRisoN,  R. :  Dietetic  deficiency  and  endocrine  activity  with  special 

reference  to  deficiency  edema.     Brit.  Med.  J.  Aug.  14,  236,  1920. 

609.  CRONHEIM:  Die  Rolle  des  Lecithins  im  Stoffwechsel  der  Erwachsene. 

Z.  physikal.  u.  diat.  Ther.  16,  262. 

610.  ROBERTSON,   T.   BRAILSFORD:  Experimental  studies  on  growth.    VI. 

The  influence  of  lecithin  upon  the  growth  of  the  white  mouse.  J. 
Bibl.  Chem.  25,  647,  1916. 

611.  MCLEAN,  HUGH:  A  simple  method  for  the  preparation  of  lecithin.     J. 

Path.  Bact.  18,  490,  1914;  The  composition  of  "lecithin"  together 
with  observations  on  the  distribution  of  phosphatides  in  the  tissues 
and  methods  for  their  extraction  and  purification.  Biochem.  J.  9, 
351,  1915 

612.  ISCOVESCO:  Les  propriety's  physiologiques  des  le"citides  du  foie.     C.  r. 

soc.  biol.  76,  74,  117,  1914.. 

613.  ROBERTSON,  T.  BRAILSFORD  :  On  the  isolation  and  properties  of  tethelin, 

the  growth-controlling  principle  of  the  anterior  lobe  of  the  pituitary 
body.  J.  Biol.  Chem.  24,  409,  1916. 

614.  COOPER,  EVELYN  ASHLEY:  The  relation  of  vitamines  to  lipoids.  Biochem. 

J.  8.  347,  1914. 

615.  SULLIVAN,  M.  X.  AND  VOEGTLIN,  CARL:    The  relation  of  lipoids  to 

vitamines.  Proc.  Amer.  Soc.  Biol.  Chem.,  J.  Biol.  'Chem;  24,  XVII, 
1916. 

616.  ARON,  HANS:  Nahrwerte.    Bio.  Z.  92,  221,  1918;  Nahrungswert  und  die 

Bedeutung  von  nahrenden  Fett.  Ibid.  103,  172,  1920. 

617.  STEPP,  W. :  Die  Lipoide  in  ihrer  Bedeutung  als  akzessorische  Nahrstoffe. 

Med.  Klin.  No.  3,  1920. 

618.  The  Same:  Ueber  Versuche  mit  lipoidfreier  Ernahrung  an  Ratten  und 

Hunden.     Z.  f.  Biol.  69,  495,  1919. 

619.  CRAMER,  W. :  Vitamines  and   lipoid  metabolism.     Proc.  Physiol.  Soc. 

May  15,  1920;  J.  Physiol.  54,  III,  1929;  Glandular  adipose  tissue; 
relation  to  other  endocrine  organs  and  to  vitamine  problem.  Brit. 
J.  Exp.  Path.  1,  184,  1920. 

620.  HOLST,  AXEL  AND  FROLICH,  TH.  :    Experimenteller  Skorbut.  Weitere 

Untersuchungen  iiber  die  Konservierung  und  Extraktion  der  spezi- 
fischen  Bestanteile  der  antiskorbutischen  Nahrungsmittel.  Z.  f. 
Hyg.  u.  Infektionsk.  75,  334,  1913. 


432  THE    VITAMINES 

621.  SMITH,  ALICE  HENDERSON:  A  historical  inquiry  into  the  efficiency  of 

lime-juice  for  the  prevention  and  cure  of  scurvy.  J.  Roy.  Army 
Med.  C.  Feb.-March,  1919;  Lane.  II,  725,  1918. 

622.  FREISE,  E. :  Der  Alkoholextrakt  aus  Vegetabilien  als  Trager  barlowheil- 

ender  Stoffe.     M.  f.  Kinderh.  12,  687,  1914. 

623  FREUDENBERG,  ERNST.  :    Beitrag  zur  Frage  des  Barlow  Schutzstoffes. 
Ibid.  13,  141,  1914-6. 

624.  HARDEN  AND  ZILVA:  Antiscorbutic  factor  in  lemon  juice.     Biochem.  J. 

12,  259,  1918. 

625.  HARDEN   AND   ROBISON,    ROBERT:    The    antiscorbutic    properties   of 

concentrated  fruit  juices.     J.  Roy.  Army  Med.  C.,  Jan.,  1919. 

626.  The  same.     The  antiscorbutic  properties  of  concentrated  fruit  juices. 

III.     Biochem.  J.  14,  171,  1920. 
626a.  VEDDER,  E.  B.:    Etiology  of  scurvy.    Milit.  Surg.49, 133, 1921. 

627.  GIVENS,  M.  H.  AND  MCCLUGAGE,  H.  B. :    Antiscorbutic  property  of 

fruits.  I.  An  experimental  study  of  dried  orange  juices.  Amer. 
J.  Dis.  Childr.  18,  30,  1919. 

628.  DUBIN,  HARRY  E.  AND  LEWI,  MAURICE  J. :  Amer.  J.  Med.  Sci.  159,  264, 

1920. 

629.  BASSETT-SMITH  :  Scurvy.    With  special  reference  to  prophylaxis  in  the 

Royal  Navy.    Lane.  May  22nd,  1102,  1920. 

630.  HARDEN  AND  ZILVA:  A  note  on  the  susceptibility  of  the  antiscorbutic 

principle  to  alkalinity.     Lane.  II,  320,  1918. 

631.  HESS  AND  UNGER:  The  scurvy  of  guinea  pigs.    III.  The  effect  of  age, 

heat,  and  reaction  on  antiscorbutic  foods.  J.  Biol  .Chem.  38,  293, 
1919. 

632.  McCLENDON,  J.  F.  AND  SHARP,  PAUL  F. :   The  hydrogen  ion  concentra- 

tion of  foods.     J.  Biol.  Chem.  38,  531,  1919. 

633.  SOMMER,  H.  H.,  AND  HART,  E.  B. :  The  effect  of  heat  on  the  citric  acid 

of  milk.    Ibid.  35,  313,  1918. 

634.  FABER,  HAROLD  K. :  Sodium  citrate  and  scurvy.    Proc.  Soc.  Exp.  Biol. 

Med.  17,  140,  1920. 

635.  FOWLER,  G.  S. :  Edinb.  Med.  J.  Jan.  2nd,  1914. 

636.  HESS,  A.  F. :  Brit.  Med.  J.  July  31st,  154,  1920. 

636a.  ZILVA,  S.  S;  AND  MIURA,  M.:    The  differential  dialysis  of  the  anti- 
neuritic  and  the  antiscorbutic  factors.    Biochem.  J.  15,  422, 1921. 

637.  HESS  AND  UNGER:  Experiments  on  antiscorbutics.     Report  of  an  anti- 

scorbutic for  intravenous  use.  Proc.  Soc.  Exp.  Biol.  Med.  15,  141, 
1918. 

638.  HESS,  A.  F. :  Infantile  scurvy.    Proc.  II,  Pan-Amer.  Sci.  Congr.  Wash. 

X,  48,  1917. 

639.  HESS,   A.    F. :   Scurvy.     Past  and  present.     Lippincott,  Philadelphia  & 

London,  1920. 

640.  CHICK,  HARRIETTS  AND  HUME,  E.  MARGARET:  Note  on  the  importance 

of  accurate  and  quantitative  measurements  in  experimental  work 
on  nutrition  and  accessory  food  factors.  J.  Biol.  Chem.  39,  203, 
1919. 


LITERATURE   TO   THE    TEXT  433 

641.  MENDEL,   L.   B. :  Food  factors  in  gastro-enterology.     Amer.   J.  Med. 

Sci.  158,  297,  1919. 

642.  RANWEZ,  F. :  Les  nouvelles  theories  sur  la  valeur  des  produits  alimen- 

taires  en  relation  avec  la  suppression  des  fraudes.  J.  pharm.  beige. 
2,  537,  1920. 

643.  COOPER,  E.  A. :  On  the  protective  and  curative  properties  of  certain 

foodstuffs  against  polyneuritis  induced  in  birds  by  a  diet  of  polished 
rice.  J.  Hyg.  12,  436,  1912;  14,  12,  1914. 

644.  CHICK,  H.  AND  DALYELL,  E. :   Skorbutgefahr  in  Wien.     Wien.  klin.  W. 

32,  1219,  1919. 

645.  CHICK,  H.  AND  HUME,  E.  M. :  Effect  of  exposure  to  temperature  at  or 

above  100°  upon  the  substance  (vitamine)  whose  deficiency  in  a 
diet  causes  polyneuritis  in  birds  and  beriberi  in  man.  Proc.  R. 
Soc.  90B,  60,  1917. 

646.  MILLER,  ELIZABETH  W. :  The  effect  of  cooking  on  the  water-soluble 

vitamine  in  carrots  and  navy  beans.     J.  Biol.  Chem.  44,  159,  1920. 

647.  WHIPPLE,  BERTHA  E. :  Water-soluble  B  in  cabbage  and  onion.    Ibid. 

44,  175,  1920. 

648.  WEILL   AND   MOURIQUAND:   Recherches   sur   le   scorbut   experimental 

Arch,  physiol.  path.  ge"n.  17,  849,  1918. 

649.  DELF,  E.  M.  AND  TOZER,  F.  M. :    Antiscorbutic  value  of  cabbage.  I. 

The  antiscorbutic  and  growth-promoting  properties  of  raw  and 
heated  cabbage.  Biochem.  J.  12,  416,  1918. 

650.  STRUDWICK,  F. :  Scurvy.    Cited  by  Med.  Res.  Comm.  No.  38,  p.  65, 1919. 

651.  DELF,  MARION  ELLEN:  Effect  of  heat  on  the  antiscorbutic  accessory 

factor  of  vegetables  and  fruit  juices.    Biochem.  J.  14,  211,  1920. 

652.  GIVENS,  MAURICE  H.  AND  MCCLUGAGE,  HARRY  B. :    Influence  of  tem- 

perature on  the  antiscorbutic  vitamine  in  tomatoes.  Proc.  Soc. 
Biol.  Chem.;  J.  Biol.  Chem.  41,  XXIV,  1920. 

653.  Rossi,  G. :  Experimental  scurvy :  the  possibility  of  sterilizing  at  high 

temperature  without  altering  the  alimentary  value  of  the  sterilized 
substances.  Arch,  fisiol.  16,  125,  1918. 

654.  DANIELS,  AMY  L.  AND  MC€LURG,  NELLIE  I. :    Influence  of  high  tem- 

peratures and  dilute  alkalies  on  the  antineuritic  property  of  food. 
J.  Biol.  Chem.  37,  201,  1919. 

655.  DANIELS  AND  HEISIG,  ESTHER:  The  acidity  of  various  sirups  used  in 

cooking.     J.  Home  Econ.  11,  193,  1919. 

656.  HESS  AND  UNGER:  The  deleterious  effect  of  the  alkalinisation  of  infants 

food.     J.  A.  M.  A.  73,  1353,  1919. 

657.  GIVENS  AND  MCCLUGAGE:  Antiscorbutic  property  of  vegetables.     II. 

An  experimental  study  of  raw  and  dried  potatoes.  J.  Biol.  Chem. 
42,  491,  1920. 

658.  HINDHEDE,  M.:  Dytidskost  samt  Kortfattet  Ernaeringslaere.    Smaas- 

kriften  ud  givne  af  det  af  Indenrigsministeriet  nedsatte  Hushold- 
nigsudvaig,  No.  10,  1918. 

659.  HILL,  J.  R. :  Food  wastage  on  potato  cooking.    Pharm.  J.  100,  149,  1918. 

660.  DENTON,  MINNA:  Change  in  food  value  of  vegetables  due  to  cooking. 

J.  Home  Econ.  11,  143,  1919. 


434  THE   VITAMINES 

661.  MURLIN,  J.  R. :  Boston  Med.  &  Surg.  J.  179,  395,  1918. 

662.  PRESCOTT,  S.  C.:  Dried  vegetables  for  army  use.    Amer.  J.  Physiol. 

49,  578,  1919. 

663.  GIVENS,  MAURICE  H.  AND  COHEN,  BARNETT:  The  antiscorbutic  property 

of  dessicated  and  cooked  vegetables.     J.  Biol  Chem.  36,  127,  1918. 
663a.  HOLST  AND  FroLicn :  Scorbutics  and  its  prophylaxis.     Norsk  Mag.    f . 
Laegerldensk.  77,  989,  1916;  J.  Trop.  Med.  Hyg.  23,  261,  1920. 

664.  GIVENS  AND  MCCLTJGAGE:  The  antiscorbutic  property  of  vegetables. 

I.  An  experimental  study  of  raw  and  dried  tomatoes.  Ibid.  37, 
253,  1919. 

665.  SHORTEN,  J.  A.  AND  RAY,  C. :    Report   on  the   antiberiberi   vitamine 

and  antiscorbutic  property  of  sun  dried  vegetables.  Proc.  Asiat. 
Soc.  Beng.  15,  226,  1919;  Ind.  J.  Med.  Res.  Spec.  No.  60,  1919. 

666.  FALK,  K.  GEORGE,  McGuiRE,  GRACE  AND  BLOUNT,  EUGENIE  :  Studies  on 

enzyme  action.  XVII.  The  oxidase,  peroxidase,  catalase  and 
amylase  of  fresh  and  dehydrated  vegetables.  J.  Biol.  Chem.  38, 
229,  1919. 

667.  BIGELOW,  W.  D. :  Problems  of  canning  operations.    Amer.  J.   Publ. 

Health  8,  212,  1918. 

668.  HESS  AND  UNGER:  Canned  tomatoes  as  an  antiscorbutic.     Proc.  Soc. 

Exp.  Biol.  Med.  16,  1,  1918. 

669.  CAMPBELL,  MABEL  E.  D.  AND  CHICK,  H. :  The  antiscorbutic  and  growth- 

promoting  value  of  canned  vegetables.     Lane.  Aug.  23,  1919. 

670.  POL,  D.  J.  HULSHOFF:  Dietetic  treatment  of  beriberi  in  the  Dutch  Indies. 

Norsk  Mag.  Laegeridenskaben.  77,  No.  1,  1916. 

671.  DELF,  E.  M.  AND  SKELTON,  R.  F. :  Effect  of  drying  on  the  antiscorbutic 

and  growth-promoting  properties  of  cabbage.  Biochem.  J.  12,  448, 
1918. 

672.  HESS  AND  UNGER:  The  scurvy  of  guinea  pigs.     II.  Experiments  on  the 

effect  of  the  addition  of  fruits  and  vegetables  to  the  dietary.  J. 
Biol.  Chem.  35,  487,  1918. 

673.  WINTON,  A.  L.,  BURNET,  W.  C.  AND  BORNMANN,  J.  H. :  Composition  of 

corn  (maize)  meal  manufactured  by  different  processes  and  the 
influence  of  composition  on  the  keeping  qualities.  Bur.  of  Chem. 
Dept.  of  Agr.  Bull.  215,  1915. 

674.  HINDHEDE,  M. :  Das  Ganzkornbrot.     Z.  physik.  u.   diatet.  Ther.   18, 

1914;  Skand.  Arch.  Physiol.  28,  165;  32,  59,  1915.  Ned.  Tijdsr.  v. 
Gen.  1,  2115,  1919. 

675.  ROHMANN,  F. :  Der  Nahrwert  des  Ganzkornbrotes.    Berl.  klin.  W.  53, 

105,  1916. 

676.  EIJKMAN,  C. :  White  bread  or  dark  bread.    Arch,  neerl.  physiol.  1,  766, 

1917. 

677.  EIJKMAN  AND  POL,  HULSHOFF:  Experiments  on  the  nutritive  value  of 

standard  white  bread  and  white  bread.  Prod.  Acad.  Sci.  Amsterd. 
21,  48,  1918. 

678.  DUTCHER,  R.  A. :  Shall  we  eat  whole  wheat  bread?  Science  47,  228,  1918. 

679.  STOKLASA,  J. :  Das  Brot  der  Zukunft.    Gust.  Fischer,  Jena  1917. 


LITERATURE   TO   THE   TEXT  435 

680.  PUGLIESE,  A. :  Valeur  nutritive  de  different  genres  de  pain.    Rev.  ge"n. 

sci.  6,  612,  1915. 

681.  VAN  LEERSUM,  R.  L.  AND  MUNK,  J. :    Nutritive  value  of  bread  from 

whole  wheat  flour.     Arch.  ne"erl.  de  physiol.  1,  446,  1917. 

682.  RUBNER,  M. :  Ganzweizenbrot.     Arch.  Physiol.  245,  1918. 

683.  v.  LIEBIG,  JUSTUS:  Ein  Mittel  zur  Verbesserung  und  Entsauerung  des 

Roggenbrotes.     Lieb.  Ann.  1854. 

684.  MAGENDIE:  Cited  according  to  Gustav  Meyer.     Z.  f.  Biol.  7,  1,  1871. 

685.  OSEKI,  S. :  Untersuchungen  iiber  qualitativ  unzureichende  Ernahrung. 

Bio.  Z.  65,  158,  1914.. 

686.  WEILL  AND  MOURIQUAND:  Note  sur  la  valeur  du  pain  de  guerre.    Soc. 

me"d.  milit.  de  la  XIV  region.    7  nov.  1915;  Acad.  de  me"d.  26  juin 
1917;  Paris  mSd.  8,  258,  1918. 

687.  SHERMAN,  H.  C.,  ROUSE,  M.  E.  ALLEN,  B.  and  WOODS  E. :    Growth  and 

reproduction  upon  simplified  food  supply.     Proc.  Soc.  Exp.  Biol. 
Med.  17,  9,  1919. 

688.  McCoLLUM  AND  DAVIS  :  The  influence  of  certain  vegetables  fat  in  growth. 

J.  Biol  Chem.  21,  179,  1915. 

689.  CHICK  AND  HUME:  Distribution  in  wheat,  rice  and  maize  grains  of  the 

substance,  the  deficiency  of  which  in  diet  causes  polyneuritis  in 
birds  and  beriberi  in  man.     Proc.  R.  Soc.  90B,  44,  1917. 

690.  VOEGTLIN,  LAKE  AND  MYERS:  The  dietary  deficiency  of  cereal  foods 

with  reference  to  their  content  in  antineuritic  vitamine.     IT.   S. 
Publ.  Health  Serv.  Repr.  33,  647,  1918. 

691.  GREIG,  E.  D.  W.  AND  CURJEL,  D.  F.:    Antiberiberi   vitamine  in  atta 

biscuits.     Ind.  J.  Med.  Res.  6,  56,  1918. 

692.  WEILL  AND  MOURIQUAND:  C.  r.  soc.  biol.  79,  194,  1916. 

693.  CHICK,  H.  AND  DELF,  ELLEN  MARION  :   The  anti-scorbutic  value  of  dry 

and  germinated  seeds.     Biochem.  J.  13,  199,  1919. 

694.  WEILL  AND  MOURIQUAND:  Des  resultats  compares  sur  la  cobaye  de 

Porge  complete,  dans  Petat  de  reposet  dans  Petat  de  germination. 
C.  r.  soc.  biol.  80,  33,  1917. 

695.  The  same  and  PERRONET,  MLLE.  :  Apparition  d'une  substance  anti- 

scorbutique  pendant  la  germination  des  grains  de  ble\     Ibid.  81, 
608,  678,  1918. 

696.  FUNK,  CASIMIR  AND  DUBIN,  HARRY  E.:  Unpublished  data. 

697.  MATTEI,  P.  D. :  Coffee  and  the  vitamines.     Policlin.  27,  1011,  1920. 

698.  WISE,  F.  B.  AND  BROOMELL,  A.  W. :  The  milling  of  rice  and  its  mechanical 

and  chemical  effect  upon  the  grain.    U.  S.  Dep.  Agr.  Bull.  330,  31  pp. 
1915. 

699.  GUARESCHI,  T.:  Use  of  rice  as  food.     Giorn.  farm.  chim.  66,  161,  1917; 

Ann.  acad.  d'agr.  Torino.  60,  41.. 

700.  SANEYOSHI:  On  kakke.     Proc.  XIII.     Intern.  Congr.  Med.  Paris.  17,  78, 

1900. 

701.  WEILL  AND  MOURIQUAND  :  Be"ribe"ri  experimental  provoque*  par  Palimen- 

tation  exclusive  par  Porge  decortique"e  ou  sterilised.     C.  r.  soc. 
biol.  78,  649,  1918. 


436  THE    VITAMINES 

702.  STEENBOCK,  H.,  KENT,  HAZEL  E.  AND  GEOSS,  E.  G. :  The  dietary  quali- 

ties of  barley.     J.  Biol.  Chem.  35,  61,  1918. 

703.  McCoLLUM,  SIMMONDS  AND  PITZ  i  The  nature  of  the  dietary  deficiencies 

of  the  wheat  embryo.     Ibid.  25,  105,  1916. 

704.  OSBORNE  AND  MENDEL:  The  nutritive  value  of  the  wheat  kernel  and  its 

milling  products.     Ibid.  37,  557,  1919. 

705.  BEZZOLA:  Beitrag  zur  Kenntnis  der  Ernahrung  mit  Mais.     I.   Einwir- 

kung  der  Maisfiitterung  auf  Meerschweinchen.     Z.  f.  Hyg.  56,  75, 
1907. 

706.  LUCKSCH:  Untersuchungen  zur  Pellagrafrage.     Z.  f.  Hyg.  58,  474,  1908. 

707.  v.  NEUSSER  :  Munch,  med.  W.  1887. 

708.  HOLST,   AXEL:  Ueber  das  Vorkommen  skorbutischer  Symptome    bei 

Pellagra  und    ihre  Erklarung.    Medicinsk  Revu's  festskrift.    Juli 
1911. 

709.  BAGLIONI.  S:  Feeding  of  corn.    Arch.  ital.  de  biol.  64,  45,  1915;  Atti 

accad.  Lincei.  24,  II,  213,  254,  1915. 

710.  DRISCOLL:  A  theory  of  the  etiology  of  pellagra.    South.  Med.  J.  6,  400, 

1913. 

711.  CLEMENTI,  A. :  Observations  of  the  effect  of  exclusive  maize  and  exclusive 

rice  diet  in  relation  to  so-called  vitamines.     Rass.  clin.  terap.  sci. 
affini  16,  121,  1917;  Arch.  farm.  sper.  2,  441,  1916. 

712.  SZALAGYI,  K.  AND  KRiwuscHA,  A.  i  Ausnutzung  des  Maises  von  Hennen, 

Enten  und  Gansen.  Bio.  Z.  88,  286,  1918. 

713.  URBEANU,  A. :  Die  Gefahr  einer    an    Kaliumverbindungen  zu  armen 

Ernahrungsweise  und  ihre  Beziehung  zu  Ernahrungskrankheiten. 
Berlin  1916. 

714.  SUAREZ,  P. :  Maisfiitterung  in  Beziehung  zu  der  Pellagrafrage.     Bio. 

Z.  77,  17,  1916. 

715.  NITZESCO,  J.  J. :  Nutrition  exclusive  des  animaux  avec  le  mais.     C.  r. 

soc.  biol  78,  222,  1915. 

716.  RONDONI,  P. :  Maize  diet  from  the  point  of  view  of  the  etiology  of  pellagra 

Speriment.  69,  723,  1915. 

717.  HOGAN,  ALBERT  G. :  The  nutritive  properties  of  corn.     J.  Biol.  Chem. 

27,  193,  1916. 

718.  MENDEL,  L.  B.  AND  FINE,  M.  S. :  The  utilization  of  the  proteins  of  the 

corn.     Ibid.  10,  1911. 

719.  OSBORNE  AND  MENDEL:  Feeding  experiments  relative  to  the  nutritive 

value  of  the  proteins  of  maize.    Proc.  Soc.  Biol.  Chem. ;  J.  Biol. 
Chem.  14,  XXXI,  1913. 

720.  BAGLIONI,  S. :  The  effects  of  corn  feeding.  VI.  Nutritive  value  of  flour 

of  wheat,  corn  and  egg  in  white  rats.     Atti  accad.  Lincei  24, 1.  1158, 
1915. 

721.  McCoLLUM  AND  SIMMONDS:  A  biological  analysis  of  pellagra  producing 

diets.     I.  The  dietary  properties  of  mixtures  of  maize  kernel  and 
bean.    J.  Biol.  Chem.  32,  29,  1017. 

722.  JOHNS,  CARL  O.,  FINKS,  A.  J.  AND  PAUL,  MABEL  S. :  Studies  in  nutrition. 

III.   The  nutritive  value  of  commercial  corn  gluten  meal.     Ibid. 
41,  391,  1920. 


LITERATURE   TO    THE    TEXT  437 

723.  OSBORNE  AND  MENDEL  :  Skimmed  milk  as  a  supplement  to  corn  in  feeding. 

Ibid.  44,  1,  1920. 

724.  WOODS,  CHARLES  D. :  Food  value  of  corn  and  corn  products.    U.  S. 

Dep.  Agr.  Bull.  298. 

725.  JURITZ:  The  chemical  composition  of  South-African  maize  and  other 

cereals.     Union  S.  Afr.  Agr.  J.  Sept.  495,  1913. 

726.  MACCRAE:  The  phosphoric  oxide  content  of  maize  flours.     J.  Hyg.  14, 

395,  1914. 

727.  POPPE,  E. :  Wanderung  der  Bestandteile  des  Maiskernes  in  Wasser  und 

in  wasserige  Losungen.     Bull.  soc.  chim.  belg.  27,  103,  1913. 

728.  McCoLLUM,   SIMMONDS  AND  PITZ  :  Dietary  deficiencies  of  the  maize 

kernel.     J.  Biol.  Chem.  28,  153,  1916. 

729.  WEILL  AND  MOURIQUAND.:  Pellagre.  Progres  me*d.  mai  1917;  C.  r.  soc. 

biol.  80,  372,  1917. 

730.  VOEGTLIN,   SULLIVAN  AND  MYERS:  Bread  as  a  food.    Change  in  its 

vitamine  content  and  nutritive  value  with  reference  to  the  occur- 
rence of  pellagra.  U.  S.  Publ.  Health  Serv.  Repr.  333,  Apr.  14th, 
1916. 

731.  HUGHES,  J.  S.:  Some  nutritive  properties  of  corn.    Kansas  Agr.  Exp. 

Sta:,  Bull.  5,  39,  1918. 

732.  HINDHEDE,  M. :  Skand.  Arch.  Physiol.  27,  277,  1912;  31,  259,  1914;  Studien 

iiber  das  Eiweissminimum.  Deutsch.  Arch.  klin.  Med.  Ill,  366, 
1913. 

733.  ABDERHALDEN,  E.  FODOR,  A.,  AND  ROSE,  C. :   Arch.  ges.  Physiol.  160, 

511,  1915. 

734.  ROSE,  MARY  S.  AND  COOPER,  LENA  F. :    The  biological  efficiency  of 

potato  nitrogen.     J.  Biol.  Chem.  30,  201,  1917. 

735.  RUBNER,  MAX:  Untersuchungen  von  Hindhede  uber  die  Verdaubarkeit 

der  Kartoffel.  Arch.  (Anat.)  Physiol.  16,  1918. 

736.  McCoLLUM,  SIMMONDS  AND  PARSONS:  The  dietary  properties  of  the 

potato.     J.  Biol.  Chem.  36,  197,  1918. 

737.  OSBORNE  AND  MENDEL:  Nutritive  factor  in  plant  tissue.     III.  Further 

observations  on  the  distribution  of  water-soluble  vitamine.  Ibid. 
41,  451,  1920. 

738.  BORUTTAU,  H. :  Ueber  das  Verhalten  von  Erganzungstoffen.     I.  Bio.  Z. 

82,  103,  1917. 

739.  AUER,  ALOYS:  Ueber  qualitativ  unzureichende  Ernahrung.    Ibid.  93, 

1,  1919. 

740.  LANE-CLAYPON,  JANET:  Value  of  boiled  milk  as  a  food  for  infants  and 

young  animals.    Rep.  Loc.  Gov.  Board  New  Ser.  No.  63,  1912. 

741.  The  same.     "The  biological  properties  of  milk"  both  of  the  human 

species,  and  of  cows,  considered  in  special  relation  to  the  feeding  of 
infants.  Ibid.  No.  76,  1913. 

742.  The  same.    Milk  and  the  Public  Health.    Longmans,  Green  &  Co., 

London. 

743.  HIROTA:  Ueber  die  durch  die  Milch,  der  an  Beriberi  leidenden  Frauen, 

verursachte  Krankheit  der  Sauglinge.  Z.  f.  inn.  Med.  No  16,  385, 
1898. 


438  THE  •  VITAMINES 

744.  ANDREWS:  Infantile  beriberi.    Philipp.  J.  Sci.  7,  67,  1912. 

745.  CARR,  R.  H.,  SPITZER,  GEORGE,  CALDWELL  R.  E.  AND  ANDERSON  C.  H. 

The  efficiency  of  certain  milk  substitutes  in  calf  feeding.  J.  Biol. 
Chem.  28,501,  1917. 

746.  HART,  E.  B.,  NELSON  V.  E.  and  PITZ,  W. :    The  synthetic  capacity  of 

the  mammary  gland.  I.  Can  this  gland  synthetize  lysine.  Ibid.  36, 
291,  1918. 

747.  McCoLLUM  AND  SIMMONDS  :  The  nursing  mother  as  a  factor  of  safety 

in  the  nutrition  of  the  young.    Amer.  J.  Physiol.  46,  275,  1918. 

748.  DRUMMOND,  J.  C. :    Some  aspects  of  infant  feeding.    Lane.   II,  482, 1918. 

749.  ECKLES,  C.  H.  AND  PALMER  L.  S. :    Influence  of  plane  of  nutrition  of 

the  cow  upon  the  composition  and  properties  of  milk  and  butter 
fat.  Influence  of  underfeeding.  Miss.  Exp.  Agr.  Sta.  Bull.  25,  107, 
1916. 

750.  ECKLES,  PALMER  AND  SWETT  W.  W. :    Factors  influencing  the  compo- 

sition of  milk.  The  influence  of  condition  at  parturition  on  com- 
position of  milk  and  butter-fat.  Ibid.  163,  33,  1919. 

751.  CHICK,  HUME  AND  SKELTON  R.  F. :   Antiscorbutic  value  of  cow  milk. 

Biochem.  J.  12,  131,  1918. 

752.  McCoLLUM,    SIMMONDS    AND   PITZ:  The  relation  of  the  unidentified 

dietary  factors,  the  fat-soluble  A  and  water-soluble  B,  of  the  diet 
to  the  growth  promoting  properties  of  milk.  J.  Biol.  Chem.  27, 
33,  1916. 

753.  HART,  STEENBOCK  AND  ELLIS  N.  R. :  Influence  of  diet  on  the  antiscor- 

butic potency  of  milk.     Ibid.  42,  383,   1920. 

754.  HESS,  UNGER  AND  SUPPLEE  G.  C. :    The  relation  of  fodder  to  the  anti- 

scorbutic potency  and  salt  content  of  milk.    Proc.  Soc.  Exp.  Biol. 

Med.  Oct.  20th,  1920. 
754a.  DUTCHER,  ECKLES,  DAHLE,  MEAD  AND  SCHAEFER:  Influence  of  diet 

of  the  cow  upon  the  nutritive  and  antiscorbutic  properties  of  cow's 

milk;  J.  Biol.  Chem.  45,  119,  1920. 
754b.  MOORE  J.  J. :  Experimental  studies  in  diet  deficiency  diseases.    Proc. 

Inst.  of  Med.  Chicago,  254.     1918. 

755.  GIBSON,  B.  B.  AND  CONCEPTION,  J.:  Philipp.  J.  Sci.     11B,  119,  1916. 

756.  CHICK,  HUME  AND  SKELTON:    The  antiscorbutic  value  of  milk  in  infant 

feeding.     Lane.    I.     1,  1918. 

757.  HESS,  A.  F. :  Scurvy  and  pasteurized  milk.    Amer.  J.  Obstetr.   and 

Dis.  Childr.  July  1916. 

758.  HESS,  A.  F. :    Infantile  Scurvy.     A  study  of  its  pathogenesis.  Amer. 

J.  Dis.    Childr.  14,  337,  1917. 

759.  HESS  AND  UNGER:  Scurvy  VIII.    Factors  affecting  the  antiscorbutic 

value  of  foods.     Ibid.     17,  221,  1919. 

760.  MILLER:   CLEVELAND.   Med.    J.   Aug.    1917. 

761.  DANIELS,  AMY  L.  AND  STUESSY  SYLVIA:    The  nutritive  value  of  boiled 

milk.    Amer.    J.    Dis.    Childr.  11,  45,  1916. 

762.  DANIELS   AND   LOUGHLIN.  :    A   deficiency    in   heat-treated   milks.     J. 

Biol.  Chem.    44,  381,  1920. 


LITERATURE   TO   THE   TEXT  439 

763.  FROLICH,    TH.  :    Experimentelle    Untersuchugen    iiber    den    infantilen 

Skorbut.     Z.  f.  Hyg.  u.  Infektionskr.     72,  155,  1912. 

764.  BARNES,   ROSAMUND  E.   AND  HUME  E.   M. :    Relative  anti-scorbutic 

value  of  fresh,  dried  and  heated  cow's  milk.  Lane.  Aug.  23rd 
1919;  Biochem.  J.  13,  306,  1919. 

765.  HART,  STEENBOCK  AND  SMITH  D.  W. :    Studies  in  experimental  scurvy. 

Effect  of  heat  on  the  antiscorbutic  properties  of  some  milk  products. 
J.  Biol.  Chem.  38,  305,  1919. 

766.  Food  Rep.  to  LOG.  Gov.  Board  No.  15,1911. 

767.  Anon:  Dried  milk  powder.   U.  S.  Publ.  Health  Rep.  No  473,  1052,  1918; 

Food  Rep.   No  24. 

768.  WINFIELD,  G. :    Some  investigations  bearing  on  the    nutritive   value 

of  dried  milk.     Loc.  Gov.   Board  Food  Rep.   24,  139,  1918. 

769.  NAISH:  PEDIATRICS.  26,  247,  1914. 

770.  PRITCHARD.     Ibid.  26,  300,  1914. 

771.  SEKINE,  HIDESABURO:    Nutritive  defect  of  condensed  milks.    J.  Tokyo 

Chem.  Soc.    41,  439,  1920. 

772.  COUTTS,  F.  J.  H. :    Upon  an  inquiry  as  to  the  dried  milks  with  special 

reference  to  their  use  in  infant  feeding.  Loc.  Gov.  Board  Food 
Rep.  24,  1,  1918. 

773.  GERSTENBERGER,  H.  J. :    Studies  in  adaptation  of  artificial  food  to 

human  milk.  Amer.  J.  Dis.  Childr.  10,  No.  4,  1915;  GERSTEN- 
BERGER, Run,  H.  O.,  BRICKMAN,  M.  J.,  LESLIE,  H.  J.,  AND  OCHSNER, 
R.  J. :  Studies  on  the  adaptation  of  an  artificial  food  to  human 
milk.  II.  A  report  of  three  years'  clinical  experience  with  the 
feeding.  S.  M.  A.  (synthetic  milk  adapted)  Ibid.  17,  1,  1919. 

774.  WASHBURN,  R.  M.  AND  JONES,  C.  H. :    Studies  of  the   values  of  diff- 

erent grades  of  milk  in  infant  feeding.  Vermont  Agr.  Exp.  Sta. 
Bull.  195,  6,  1916. 

775.  MATTILL,  H.  A.  AND  CONKLIN,  RUTH  E. :      The  nutritive  properties 

of  milk,  with  special  reference  to  reproduction  in  the  albino  rat. 
J.  Biol.  Chem.  ~44,  137,  1920. 

776.  OSBORNE  AND  MENDEL:    Nutritive  factors  in  animal  tissues.  I.  J.  Biol. 

Chem.   32,  309,  1917. 

777.  OSBORNE  AND  MENDEL:    Nutritive  factors  in  animal  tissues  II.  Ibid. 

34,  17,  1918;  Proc.  Soc.  Exp  Biol.  Med.  15,  71,  1918. 

778.  COLE,  S.  W. :    Report  on  a  dried  meat  powder.    Food   (War)    Comm. 

R.  Soc  Lond.  6,  1917. 

779.  NANSEN,   FRIDJOF.  :    Voyage  on  Fram. 

780.  STEFANSSON,  V.:  J.  A.  M.  A..  71,  1715,  1918. 

781.  CURRAN:  DUBLIN  J.  Med.  Sci.  7,  83,  1847. 

782.  WILLCOX,  W.  H. :  Rations  in  relation  to  disease  in  Mesopotamia.     Lane. 

II.   677,  1917. 

783.  DUTCHER,  R.  A.,  PIERSON  EDITH  M.  AND  BIESTER,  ALICE:  The  anti- 

scorbutic properties  of  raw  beef.  Science.  50,  184,  1919;  J.  Biol. 
Chem.  42,  301,  1920. 

784.  GIVENS,  M.  H.  AND  MCCLUGAGE  H.  B. :  The  antiscorbutic  property 

of  dehydrated  meat.    Science  51,  273,  1920. 


440  THE    VITAMINES 

785.  McCoLLUM  AND  DAVIS  :  The  influence  of  certain  vegetable   fats  on 

growth.     J.  Biol.  Chem.  21,  179,  1915. 

786.  WHEELER,  RUTH  AND  BIESTER,  ALICE  :  A  sfcady  of  the  nutritive  value 

of  some  proprietery  infant  foods.  After.  J.  Dis.  Childr.  7,  169, 
1914. 

787.  WHEELER,  R. :  A  study  of  the  nutritive  value  of  some  proprietary  infant 

foods.   II.     As  food  modifiers.     Ibid.  9,  300,  1915. 

788.  OSBORNE  AND  MENDEL:  The  vitamines  in  green  foods.     J.  Biol.  Chem. 

37,  187,  1919. 

789.  ABDERHALDEN,  E. :  Studien  iiber  den  Einfluss  der  Art  der  Nahrung 

auf  das  Wohlbefinden  des  einzelnen  Individuums,  seine  Lebens- 
dauer,  seine  Fortpflanzungsfahigkeit  und  das  Schicksal  der  Nach- 
kommen.  Arch.  ges.  Physiol.  175,  187,  1919. 

790.  SWOBODA,  FREDERICK  H. :  A  quantitative  method  for  the  determina- 

tion of  vitamine  in  glandular  and  other  tissue.  J.  Biol.  Chem. 
44,  531,  1920. 

791.  HINDHEDE,  M. :  Ernahrungsversuche  mit  grob  zerquetschtem  Weizen. 

Skand.  Arch.  Physiol.  33,  263,  1916. 

792.  HESS,  A.  F. :  The  therapeutic  effect  of  wheat  embryo  and  of  yeast  in 

infantile  scurvy.  Proc.  Soc.  Exp.  Biol.  Med.  13,  145,  1916;  Amer. 
J.  Dis.  Childr.  13,  98,  1917. 

793.  SHERMAN,   H.   C. :  Protein  requirement  of  maintenance  in  man  and 

nutritive  efficiency  of  bread  protein.    J.  Biol.  Chem.  41,  97,  1920. 

794.  OSBORNE  AND  MENDEL:  The  food  value  of  soy  bean  products.     Proc. 

Soc.  Exp.  Biol.  Med.  14,  174,  1917. 

795.  DANIELS,  AMY  L.  AND  NICHOLS,  NELL  B. :  The  nutritive  value  of  the 

soy  bean.     J.  Biol.  Chem.  32,  91,  1917. 

796.  OSBORNE  AND  MENDEL.  :  The  use  of  soy  bean  as  food.     Ibid.  32,  369, 

1917. 

797.  WALLIS,  K.  L.  MACKENZIE:  Food  value  of  the  ground  nut   (arachis) 

Ind.  J.  Med.  Res.  6,  45,  1918. 

798.  DANIELS,    AMY   L.  AND  LOUGHLIN,    ROSEMARY:  Feeding   experiments 

with  peanuts.     J.   Biol.   Chem.  33,  295,  1918. 

799.  GREIG,  E.  D.  W. :  Report  on  antiberiberi  vitamine  content  of  ground 

nut  (peanut)  meal  biscuits.     Ind.  J.  Med.  Res.  6,  143,  1919. 

800.  McCoLLUM,  SIMMONDS  AND  PITZ  :  The  dietary  deficiencies  of  the  white 

bean,  Phaseolus  vulgaris.     J.  Biol.  Chem.  29,  521,  1917. 

801.  RICHARDSON,  ANNA  AND  GREEN,  HELEN  S. :  Nutrition  investigations 

upon  cottonseed  meal.  III.  Cottonseed  flour.  The  nature  of 
its  growth-promoting  substances,  and  a  study  in  protein  minimum. 
J.  Biol.  Chem.  31,  379,  1917. 

802.  RICHARDSON.    I  &  II.     Ibid.,  25,  307,  1916;  30,  243,  1917. 

803.  HOLMES,  ARTHUR  D. :  Digestibility  of  steam  cooked  soy  bean  and  pea- 

nuts.    J.  A.  M.  A.  74,  798,  1920. 

804.  JOHNS,  C.    O. :  The  proteins  of  the  peanut.     Cotton  Oil  Press.    2,  No. 

12,  41,  1919. 

805.  McCoLLUM,  SIMMONDS  AND  PARSONS:  The  dietary  properties  of  the 

pea  (Vicia  sativa)    J.  Biol.  Chem.  37,  287,  1919. 


LITERATURE    TO    THE    TEXT  441 

806.  WILTSHIRE,  H.  W. :  The  value  of  germinated  beans  in  the  treatment 

of  scurvy.     Lane.  II.  811,  1918. 

807.  WELLS,  C.  A.  AND  EWING  P.  V:  Cottonseed  meal  as  an  incomplete  food. 

J.  Biol.  Chem.  27,  15,  1916. 

808.  OSBORNE  AND  MENDEL:  The  nutritive  value  of  some  cottonseed  products 

in  growth.     Proc.   Soc.   Exp.   Biol.   Med.  13,  147,  1916. 

809.  STEENBOCK   AND    GROSS:  Fat-soluble  vitamine.    IV.     The   fat-soluble 

vitamine  content  of  green  plant  tissues  together  with  some  obser- 
vations on  their  water-soluble  vitamine  content.  J.  Biol.  Chem.  41, 
149,  1920. 

810.  STEENBOCK  AND  GROSS:  Fat-soluble  vitamine.  II.    The  fat-soluble  vita- 

mine  content  of  roots,  together  with  some  observations  on  their 
water-soluble  vitamine  content.  Ibid.,  40,  501,  1919. 

811.  OSBORNE    AND  MENDEL:  Nutritive   factors  in  plant  tissues.     II.    The 

distribution  of  water-soluble  vitamine.     Ibid.,  39,  29,  1919. 

812.  CHICK,  H.  AND  RHODES,  MABEL:  The  antiscorbutic  value  of  the  raw 

juice  of  root  vegetables  with  a  view  of  their  adoption  as  an  adjunct 
to  the  dietary  of  infants.  Lane.  II,  774,  1918. 

813.  JOHNS,  C.  O.,  FINKS,  A.  J.  AND  PAUL,  M.  S.:  Studies  in  nutrition.  I. 

The  nutritive  value  of  coconut  globulin  and  coconut  press  cake. 
J.  Biol.  Chem.  37,  497,  1919. 

814.  JANSEN,  B.  C.  P.:  Fat-soluble  vitamine  in  coconut  oil.    Med.  v.  d. 

Burg.  Gen.  Dienst.  Ned.  Indie.  No  6,  78,  1918;  Gen.  Tijdsr.  v. 
Ned.  Indie.  58,  173. 

815.  PICKARD,  GLENN  N. :  The  manufacture  of  nut  margarine.    Amer.  Food 

J.  13,  16,  1918. 

816.  CAJORI,  F.  A.:  Some  nutritive  properties  of  nuts;  their  proteins  and 

content  of  water-soluble  vitamine.  Proc.  Soc.  Exp.  Biol.  Med. 
17,  78,  1920;  J.  Biol.  Chem.  43,  583,  1920. 

817.  MCLEAN,  STAFFORD:  Case  report  of  scurvy  with  a  summary  of  fifty 

other  cases.     Arch.  Pediatr.  35,  477,  1918. 

818.  CHICK,   HUME  AND  SKELTON:  The    relative  content  of  antiscorbutic 

principle  in  limes  and  lemons.     Lane.   II,  735,  1918. 

819.  HESS,  A.  F. :  The  role  of  antiscorbutics  in  our  dietary.     J.  A.  M.  A. 

71,941,  1918. 

820.  LEWIS,  HOWARD  B. :  The  antiscorbutic  value  of  the  banana.    J.  Biol. 

Chem.  40,  91,  1919. 

821.  CHICK,  HUME  AND  SKELTON.  The  antiscorbutic  value  of  some  Indian 

dried  fruits.     Lane.  Aug.  23rd  1919. 

822.  JANSEN,  B.  C.  P. :  Coconut  press  cake  as  food  for  man.    Med.  v.  d.  Burg 

Gen.  Dienst.  Ned.  Indie  No.  1,  3,  1920. 

823.  VAN  DER  WTELEN,  P..  The  preparation  of  malt  extract  containing  vita- 

mine.  Pharm  Weekblad.  52,  673,  1915. 

824.  HARDEN  AND  ZILVA:  Investigation  of  beer  for  antineuritic  and  anti- 

scorbutic potency.     J.  Inst.  Brewing.  24,  197,  1918. 

825.  BUTCHER,  A.   R. :  Vitamine  studies.   III.    Observations  on  the  cura- 

tive properties  of  honey,  nectar  and  corn  pollen  in  avian  polyneu- 
ritis.  J.  Biol.  Chem.  36?  551,  1918. 


442  THE    VITAMINES 

826.  CHICK   AND   HTJME:  Distribution   among  foodstuffs    (especially   those 

suitable  for  the  rationing  of  the  armies)  of  the  substances  required 
for  the  prevention  of  beriberi  and  scurvy.  J.  R.  Army  Med.  C. 
29,  121,  1917. 

827.  FABER,  HAROLD  K. :  A  study  on  the  antiscorbutic  value  of  honey.   J. 

Biol.  Chem.  43,  113,  1920. 

828.  McCLENDON,  J.  F.  AND  COLE,  W.  C.  C.  i  The  antiscorbutic  properties 

of  green  malt.  Proc.  Amer.  Physiol.  Soc.  Amer.  J.  Physiol.  49, 
145,  1919. 

829.  GERSTENBERGER,  HENRY  J. :  Has  malt-soup  extract  an  antiscorbutic 

value?  Amer.  Pediatr.  Soc.  May  31- June  2nd  1920;  J.  A.  M.  A.  75, 
99,  1920. 

830.  OSBORNE  AND  MENDEL:  The  influence  of  beef  fat  on  growth.     Proc. 

Soc.  Exp.  Biol.  Med.  12,  92,  1915. 

831.  EDDY,  WALTER  H. :  The  isolation  of  a  growth-producing  substance  from 

sheep  pancreas.    J.  Biol.  Chem.  27,  113,  1916. 

832.  SEKINE,  HIDESABURO:  Vitamine,  fat-soluble  A,  in  fish  oils.     J.  Tokyo 

Chem.  Soc.  41,  426,  1920. 

833.  EMMETT,  A.  D.  ANT>LUROS,  G.  O.:  The  absence  of  fat-soluble  A  vitamine 

in  certain  ductless  glands.     J.  Biol.. Chem.  38,  441,  1919. 

834.  HALLIBURTON,  W.  D.  AND  DRTJMMOND,  J.  C. :  The  nutritive  value  of 

margarine  and  butter  substitutes  with  reference  to  their  content 
of  the  fat-soluble  accessory  growth  substance.  J.  Physiol.  51, 
235,  1917. 

835.  DRUMMOND,   J.   C:  The  nutritive  value  of  certain  fish.     J.   Physiol. 

52,  95,  1918. 

836.  SUZUKI,  UMETARO,  OKUDA,  SUZURU,  OKIMOTO,  TAMARO,  AND  NAGASAWA, 

TAMARY:  Nutritive  value  of  various  proteins.  I.  Nutritive 
value  of  muscle  proteins  of  marine  animals.  J.  Tokyo  Chem. 
Soc.  40,  385,  1919. 

837.  McCoLLUM  AND  DAVIS  :  Nutrition  with  purified  food  substances.     J. 

Biol.  Chem.  20,  641,  1915. 

838.  VEDDER,  EDWARD  B. :  Beriberi.    John  Bale  Sons  and  Danielson,  Lon- 

don 1913. 

839.  CASTELLANI  AND   CHALMERS:  Manual  of  Tropical  Medicine.    3d  Ed. 

840.  SCHILLING,  V. :  TropischeStoffwechselkrankheiten.    Beriberi  Spezielle 

Path.  u.   Ther.   d.   inn.  Krankh.  Kraus-Brugsch.     II  a,  949,  1915. 

841.  FINDLAY:  Beriberi.  Practitioner.  98,  69,  1917. 

842.  BALZ  AND  MIURA:    Beriberi.  Mense's  Handbuch  der  Tropenkrankhei- 

ten.     II.     Bd.     S.  140. 

843.  FRASER,  HENRY:  Rep.  from  the  Inst.  of  Med.  Res.  Fed.  Malay  States 

Apr.  Ist-Sept.  30th,  1912. 

844.  SIMPSON,  K. :  A  note  on  the  enviromental  factor  in  the  causation  of 

beriberi.    Lane.  197,  1027,  1919. 

845.  HEPBURN,  H.  H. :  Analysis  of  early  cases  of  beriberi.    Br.  Med.   J. 

Apr.  3rd,  466,  1920. 

846.  MULVANY:    Beriberi.    Ind.  Med.  Gaz.  52,98,  1917. 


LITERATURE   TO   THE   TEXT  443 

847.  CHEVALIER:    Report  on  beriberi  in  Serenli.    East  Africa  Protect.     Trop. 

Dis.  Bull.  1,  487,  1913. 

848.  DUBOIS  AND  CORIN:  Rapport  sur    une    petite    Epidemic  de  beriberi 

a  Bokala  (Congo  Beige)  Bull.   soc.  path.  exot.  7,  402,  1914. 

849.  LOVELACE,  CARL.  The  etiology  of  beriberi,  J.  A.  M.  A.  59,  2134,  1912. 

850.  WOLCOTT,  A.  M:   J.  A.  M.  A.  65,  2145,  1915. 

851.  FRAGA,  C.:  Brazil  Med.  31,  20th-27th  Jan.;  Feb.  3rd,  1917. 

852.  RIDDELL,  J.  D.,  SMITH,  CHAS.  H.  AND  IGRAVIDEZ  P.  G. :  Beriberi  at 

U.  S.  Army  Base  Hospital,  San  Juan,  Porto  Rico.    J.  A.  M.  A.  72, 
569,  1919. 

853.  LITTLE  :  Beriberi  caused  by  white  flour.  J.  A.  M.  A.  58,  2029,  1912. 

854.  PARKER:  A  report  on  beriberi  in  the   county  jail  at  Elizabeth,  N.  J. 

Publ.  Health  Rep.  29,  339,  1914. 

855.  REED:  Beriberi.  California  State  J.  Med.  15,  158,  1917. 

856.  TRAVIS:    Beriberi.    Kentucky  Med.  J.  15,  476,  1917. 

857.  CHANTEMESSE  AND  RAMOND:  Cited  by  Schiiffner.  M.  med.  W.  642,  1913. 

858.  SCHUFFNER,  W. :  1st  Beriberi  eine  in  Europa  endemische  Krankheit. 

M.  med.  W.  No.    12,  642,  1913. 

859.  MARTINEZ,   F.   F. :  Beriberi  en  Espagne  et  Portugal.     Paris  me"d.  9, 

54,  1919;  Arch.  med.  beige.  73,  299,  1920. 

860.  SICARD,  ROGER  AND  RIMBAUD:  Paris  me"d.  1  dec.  1917. 

861.  ROGER,  H:  Le  be'ribe'ri  en  France.    Med.  1,  151,  1919. 

862.  LEGGATE,  A.  R. :  Observations  on  beriberi  among  the  Chinese  in  France. 

Edinb.  Med.  J.  24.  32,  1920. 

863.  MASSALONGO:  Boll,  delle  cliniche.  31,  marz.  1917. 

864.  WILLCOX,  W.  H. :  Beri-beri,  with  special  reference  to  prophylaxis  and 

treatment.     Lane.  190,  553,  1916. 

865.  WILLCOX,   W.  H.   Treatment    and   management  of    diseases    due    to 

deficiency  of  diet:  scurvy  and  beriberi.     Brit.  Med.  J.  I,  73,  1920. 

866.  BRADDON,    LEONARD:  Investigation   as   to   occurrence   of   beriberi   in 

Mediterranean  area.    Med.  Res.  Comm.  Rep.  No  38,  p.  55. 

867.  HEHIR:    Beri-beri.    Proc.    Asiat.    Soc.    Bengal  15,  212,  1919;  Mesopot. 

Comm.  Rep.  Appendix.  Ill,  1917. 

868.  SPRAWSON,  C.  A.:  Beri-beri  in  Mesopotamian  Force.  Quart.  J.  Med. 

13,  337,  1920. 

869.  STRONG,  RICHARD  P.  AND  CROWELL,  B.  C. :  The  etiology  of  beriberi. 

Philipp.  J.  Sci.  7B,  271,  1912. 

870.  CASPARI   AND  MOSZKOWSKI:  Weiteres   zur  Beriberifrage.    Berl.   klin. 

W.  50,  1515,  1913;  D.  med.  W.  39,  1479,  1529,  1913. 

871.  FRAGA,  C.:  Beriberi.     Brazil  med.  33,  49,  1919. 

872.  CHAMBERLAIN,  WESTON  P.:  Prevention  of  beriberi  among  " Philippine 

Scouts,"  by  means  of  modification  in  the  diet.    J.  A.  M.  A.  64, 
1215,  1915. 

873.  KATO,  S.  AND  YAMADA  S. :  Arrythmia  with  beriberi.    Mitt.  d.  med.  Fak. 

Tokyo  Univers.  19,  229,  1918. 

874.  DOYLE.:  The  clinical  manifestations  of  beriberi.  N.  Y.  Med.  J.  Apr. 

8th,  1916. 


444  THE    VITAMINES 

875.  CHUN:    Beriberi.     Nation.  Med.  J.  Shanghai.     3,  113,  1917. 

876.  YOSHIKAWA,  I,  YANO  K,  AND  NEMOTO,  T. :  Studies  of  the  blood  in  beri- 

beri.    Arch.  Internal  Med.  20,  103,  1917. 

877.  JIDA,  H. :  Examination  of  the  cerebrospinal  fluid  in  beriberi.     Chagai 

IjiShimpo  895,812,  1917. 

878.  SICARD,  J.  A.  AND  ROGER  H. :  Le  liquid  cerebrospinal  dans  le  beriberi. 

Bull,  et  mem.  soc.  med.  hopitaux  de  Paris.  34,  137,  1918. 

879.  GREIG:  Epidemic    dropsy.     Scient.  memoirs  by  officers  of  the  medical 

and  sanit.  dep.  of  the  Gov.  of  India.     New  Ser.  No.  45,  1911. 

880.  Greig:    Note  on  an  investigation  on  the  causation  of  epidemic  scurvy 

in  Calcutta.     Trans.  Soc.  Trop.  Med.  Hyg.  5,  106,  1912. 

881.  LAGANE:  Le  nourisson.  No.  5,  Sept.  1913. 

882.  WEILL,  E.  AND  MOURIQUAND  G. :  La  notion  de  la  "carence"  en  patho- 

logic infantile.     Paris  med.  3  fevr.  1917. 

883.  WALSHE,  F.  M.  R. :  "Food  deficiency  or  "vitamine."  theory  in    its 

application  to  infantile  beri-beri.     Br.  J.  Childr.  Dis.  15,  258,  1918. 

884.  OGATA,  J.  AND  JIZUKI  S.:  Ueber  den  Einfluss  der  mutterlichen  Kakke 

auf  den  Fotus.     Beitr.  z.  Geburtsh  u.  Gynak.  17,  196,  1912-13. 

885.  ROXAS,  MANUEL:  Cited  according  to  McCollum.     Newest  Knowledge 

of  Nutrition,   p.  127. 
'886.  DtiRCK,  H. :  Pathologische  Anatomie  der  Beriberi.     Jena  1908. 

887.  BILZ:  Z.  f.  klin.  Med.  4,  616,  1882. 

888.  ELLIS,  W.  G. :    Lane.  II,  985,  1898. 

889.  RUMPF  AND  LUCE:  Deutsch.  Z.  f.   Nervenheilk.  18,  63,  1900. 

890.  MOTT:    The  histological  changes  in  the  nervous  system  of  Dr.  Box  case 

of  pellagra,  compared  with  changes  found  in  a  case  of  pellagra 
dying  in  Abbasieh  Asylum,  Cairo.  Trans.  Soc.  Trop.  Med.  Hyg. 
6,  156,  1913. 

891.  SUZUKI,  T.  The  amount  of  sugar  in  the  blood  of  nurslings  suffering 

from  beriberi.  Tokyo  Igakukai  Zasshi.  (19)  30,  29,  1916;  Jikwa 
Zasshi  No.  199,  41,  1916. 

892.  DE  LANGEN,  SCHUT,  WECHUIZEN  AND  ALTING:  Fat  and  lipoid  content 

of  blood  in  the  tropics.  Med.  Gen.  Labor,  te  Welteverden,  Java, 
No  2-3,  44,  1919. 

893.  ARIMA,  E. :  The  non-protein  nitrogen  blood  content  in  cases  of  beriberi. 

Tokyo  Igakukai  Zasshi.  30,  1,  1916. 

894.  SUZUKI,  T. :  Sugar  content  of  cerebrospinal  fluid  in  various  diseases  of 

children.     Tokyo  Igaku  Zasshi.  30,  231,  1916. 

895.  SUZUKI:  Amino-acid  content   of  the  urine  in  beriberi.    Kyoto  Igaku 

Zasshi.  14,  249,  1917. 

896.  PAGNIEZ    AND    VALLERY-RADOT:  Beriberi.     Ann.  de  me'd.  4,  45,  1917. 

897.  ONO,  S. :  Adrenaline  content  of  the  suprarenal  glands  of  cadavers  of  pa- 

tients dying  from  beriberi.  Jikwa  Zasshi.  Dec.  20th,  1014,  1916; 
Tokyo  Igakukai  Zasshi  31,  1,  1917. 

898.  SALEEBY,  N.  M.:  Treatment  of  human  beriberi  with  autolyzed  yeast 

extract.     Philipp.   J.  Sci.  14,  11,  1919. 


LITERATURE    TO    THE   TEXT  445 

899.  TEL  ROSARIO,  M.  V.,  AND  MARATHON,  JoAQUiN. :  Physico-chemical  valua- 

tion of  tiki-tiki  (rice  polishings)  extract.  Philipp.  J.  Sci.  15, 
221,  1919. 

900.  ALBERT,    JOSE:  Treatment  of  infantile  beriberi  with  the  extract  of 

tiqui-tiqui.     Ibid.  10B,  81,  1915. 

901.  Cox,  A.  J. :  Annual  Rep.  Philipp.  Bur.  of  Sci.  1918. 

902.  NOCHT:  Festschrift  zum  60  Geburtstag  von  R.  Koch.  p.  203,  1903. 

903.  HOLST   AND   FROLICH.  :  Experimentelle   Unterschungen   liber   die   Ur- 

sache  des  Skorbuts.  Verh.  d.  6  Norddeutsch.  Kongr.  f.  inn.  Med. 
p.  328,  1909. 

904.  HOLST,    AXEL:  The    etiology    of    beriberi.     Trans.   Soc.   Trop.   Med. 

Hyg.  5,  76,  1911. 

905.  DELPECH:  Le  scorbut  pendant  le  siege  de   Paris.    Ann  d'  hyg.  publ. 

et  meU  le"g.  II  ser.  janv.  35,  1871. 

906.  BUCQUOY:  Union  m6d.  Sept.  Oct.   1871. 

907.  GARCIA:  Case  of  associated  scurvy  and  beriberi.     Siglomed.  67, 361, 1920. 

908.  SCHERER:  Ueber  Skorbut  in  Deutsch  Siidwestafrika.    Arch.   f.  Schiffs- 

u.  Tropenhyg.  17,  191,  1913. 

909.  GOUZIEN:  Le    beriberi    au    Tonkin.    Ann.    d'hyg.,     et    m6d.     colon. 

15,  445,  1912. 

910.  FLEMING,  MACAULAY  AND  CLARK:  Report  on  the  prevalence  and  pre- 

vention of  scurvy  and  pneumonia  in  Southern  Rhodesia  among 
native  laborers,  1910. 

911.  ORENSTEIN,  A.  J. :  Annual  Rep.  Sanit.  Rand  Mines  Ltd.  1915-16. 

912.  DARLING:  The  pathological  affinities  of  beriberi  and  scurvy.    J.  A. 

M.  A.  63,  1290,  1914. 
913  DARLING:  Trop.  Dis.  Bull.  5,  1915. 

914.  SCHRODER:  Ein  Beitrag  zu  Geschichte  des  Skorbuts.    Arch,  f .  Schiffs- 

u.   Tropenhyg.  17,  263,  1913. 

915.  SCHELENZ:  Scharbock:  Ibid.  16,  113,  1912. 

916.  HIRSCH,  AUGUST:  Handbook  of  geographical  and  historical  pathology. 

London  1885.     New  Sydenham  Society. 

917.  AUTRAN:  Essai  historique  sur  le  scorbut  et  le  b£rib6ri.    These  de  Lyon. 

1916. 

918.  HESS,  ALFRED  F. :     Scurvy,  Past  and  Present.     J.   B.  Lippincott  Co. 

Philadelphia   1920. 

919.  Captain  Cook's  Voyages.     Everyman  Ed. 

920.  ELAINE,  SIR  GILBERT:  Observations  on  the  diseases  incident  to  seamen. 

London  J.  Murray  1789. 

921.  BUDD:  Scurvy.   Tweedie's  System  of  Practical  Medicine.     London  1840. 

922.  MACNAB:  Quart.  J.  Calcutta  Med.  Phys.  Soc.  1,  306,  1837. 

923.  Medical  and  Surgical  History  of  the  War  of  the  Rebellion.  1888  Wash. 

I,  Part  3,  Chapter  8. 

924.  GRENET,  A.  L.  Z. :  Le  scorbut  au  Fort  de  Bicetre  pendant  le  sie"ge  de 

Paris  par  les  prussiens.  L'hiver  de  1870-71.  Ann  d'hyg.  publ., 
36,  II  ser.  1871;  Hayem,  M.  G. :  Gaz.  m6d.  No.  13, 186,  1871;  Lasegne, 
Ch.  and  Legroux,  A.:  Arch.  gen.  de  meU  2,  680,  1871;  Lev6n,  M.: 
Gaz.  med.  de  Paris  p.  469,  1871;  Dechambre:  Ibid.  p.  147,  1871. 


446  THE    VITAMINES 

925.  SATO,  T.,  AND  NAMBU,  K. :  Zur  Pathologic  and  Anatomic  des  Skorbuts 

Virch.  Arch.  194,  151,  1908. 

926.  BLAU:  Der  Skorbut  in  der  russischen  ^.rmee   und  Marine.     Deutsch. 

militararztl.   Z.  38,  657,  1909. 

927.MACPHERSON,  SIR  W.  G. :  R.  Soc.  Med.  May  10th  1920;    Lane.     May 
22nd,  p.  1112,  1920. 

928.  RAE:  Rep.  to  Scurvy  Commission  in  1876  about  the  Hudson  Bay  Co. 

cited  by  Med.  Res.  Comm.  No.  38,  p.  63,  1919. 

929.  LANCERAUX:  Le  scorbut  des  prisons  du  department  de  la  Seine.     Ann. 

d'hyg.  publ.  et  med.  leg.  13,  III,  Ser.  1885. 

930.  BERTHENSON:  Zur  Statistik  und  Aetiologie  des   Skorbuts.  Die  Skor- 

butepidemie  von  1889  nach  Beobachtungen  in  St.  Petersburger  Mili- 
tarhospital.    Arch.  f.  klin.  Med.  49,  127,  1892. 

931.  MULLER:  Mtinch.  med.  W.  No.  35,  1894,  1911. 

932.  TAUSSIG,    SIGISMUND:  Zur  Epidemiologie   des   Skorbuts,  Militarmed. 

und  arztl.  Kriegswiss.     Heft  IV,  1913,  Safar  Wien. 

933.  PICKENS,  R.  M.:  A  death  from  scurvy.    Lane.  II.  21,1917. 

934.  HARLAN,  G.  P. :  Land  scurvy  in  England.    Brit.  Med.  J.  II,  46,  1917. 

935.  Report  on  the  Health  of  the  City  of  Manchester  49,  1917. 

936.  Edit.  Recent  cases  of  scurvy  in  Glasgow.    Brit.  Med.  J.  II,  Jly  7,  1917. 

937.  Memorandum  on  food  and  scurvy.    Food  (War)  Comm.  of  the  Roy. 

Soc.   Lane.     Nov.  40,  p.  756,  1919. 

938.  LIND,  W.  A.  T. :  Outbreak  of  scorbutus   at  Kew  Victoria.  Med.   J.  of 

Austr.    2,  107,  1919, 

939.  HOPKINS,  G.  R. :  The  etiology  of  scurvy.  J.  A.  M.  A.  69,  1641,  1917. 

940.  TURNER:  Scurvy.  Brit.  Med.  J.  II.  July  14,  33,  1917. 

941.  Trop.  Dis.  Bull.  12,  257,  1918. 

942.  HILL,  LEONARD:  Cited  by  Med.  Res.  Comm.  No.  38,  p.  65. 

943.  HORSCHELMANN,  E. :  Zur  Klinik  des  Skorbuts  in  der  russischen  Armee. 

Deutsch.  med.  W.    52,  1617,  1917. 

944.  DISQUE:  Entstehung   und  Verlauf  des  Skorbuts  im  Jahre  1916  unter 

den  deutsch-osterr.  Kriegsgefangenen  in  Taschkent  (Turkestan) 
Med.  Klin.  No  1,  1918. 

945.  BORICH,  R. :  Skorbut.  D.  Arch.  f.  klin.  Med.     130,  151,  1919. 

946.  HARVIER,  P.:  Epidemic  de  scorbut.  Paris  med.  7,  394,  1917. 

947.  BENOIT,  A. :  Une  epidemic  de  scorbut.     Ibid.    9,  469,  1919. 

948.  MOURIQUAND,  G. :  Role  de  Paliment  frais  dans  la  nutrition.     Imprim. 

milit.  Paris  1915;  La  dietetique  sur  le  front.  Arch,  de  m6d.  et 
de  pharm.  milit.  sept.  1915.  L'alimentation  aux  armees.  Lyon  me*d. 
oct.  1915. 

949.  FERRARI:  Gazz.  d.  osp.  38,  778,  1917. 

950.  RAMOINO:  Policlin.  24,  616,  1917. 

951.  GINGUI,  F. :  Rif.  med.  34,  22,  1918. 

952.  VANUTELLI,  F. :  Un  epidemia  di  porpora  infettiva  con   manifestazio- 

ne    emorragico-scorbutiche    tra    le    truppe    combattenti.     Ibid 
24,  873,  1919. 

953.  VALLARDI,  C.:  Rif.  med.  34,  793,  1918. 


LITERATURE   TO    THE    TEXT  447 

954.  MUCH,  HANS:  Ueber  den  Skorbut.    Munch,  med.  W.  64,  854, 1917;  Much 

and  Baumbach,  K. :  Ibid.  No.  26,  1917;  Saxl,  P.  and  Melka,  J. : 
Ueber  den  Skorbut  und  seine  Beziehungen  zu  den  hamorrhagischen 
Diathesen.  Med.  Klin.  No.  37,  986,  1917;  Feig,  S.:  Beobachtungen 
iiber  Skorbut  in  Kriege.  Ibid.  No.  31,  832,  1917;  Schneider,  E. : 
Skorbut  in  Felde.  Munch.  Med.  W.  No.  44,  1939,  1917;  Hertz,  A.: 
Wien.  klin.  W.  No.  22,  1917;  v.  Jaksch,  R. :  Z.  f.  inn.  Med.  No. 
32-33, 1917;  Hannemann,  K. :  Munch,  med.  W.  p.  665, 1918;  Morawitz : 
Ibid.  65,  349,  1918;  Tlichler:  Med.  Klin.  No.  5,  112,  1918;  Blatt, 
N.:  Wien.  klin.  W.  31,  942,  1918;  Arneth:  D.  med.  W.  64,  509,  1918. 

955.  SCHBEIBER,  G. :  Scorbut  et  dysenteric.  Paris  med.  p.  508,  1918. 

956.  SPEYER:  Wadenabszesse  bei  Skorbut.  D.  med.  W.  544,  626,  1918. 

957.  LOBMEYER,  G. :  Kriegschirurgische  Bedeutung  des  Skorbuts.  D.  med. 

W.  44,  557,  1918. 

958.  Edit.  Fixed  diet  in  prisons.     Trop.  Dis.  Bull.  4,  446,  1914. 

959.  DYKE,    H.   W. :  Scurvy.     Lane.    II,  513,  1917. 

960.  COMRIE,  J.  D. :  Scurvy  in  North  Russia.  Edinb.  Med.  J.  24,  207,  1920. 

961.  STEVENSON:  J.  Roy.  Army  Med.  C.  35,  218,  1920. 

962.  CHICK,  H.  AND  DALYELL,  E.  J. :  Influence  of  overcooking  vegetables 

in  causing  scurvy  among  children.  Brit.  Med.  J.  Oct.  9,  546,  1920. 
Epidemic  des  Skorbuts  bei  Schulkindern.  Z.  f .  Kinderh.  26,  257,  1920. 

963.  TOBLER,  W. :  Skorbut  im  Kindesalter.  Z.  f.  Kinderh.  18,  63,  1918. 

964.  MULLER,  ERICH:  Ueber  ein  gehauftes  Auftreten  von  Skorbut  beiKindern. 

Berl.  klin.  W.  55,  1024,  1918. 

965.  WEILL  AND  DUFOURT  A.:  Le  scorbut  tardif  chez  les  enfants.  Arch. 

me*d.  d.  enfants.  22,  561,  1919. 

966.  MORSE,  J.  L. :  Resume  of  literature  of  infantile  scurvy  during  the  past 

5  years.  Boston  Med.  Surg.  J.  178,  160,  1918;  for  1918-9.  Ibid. 
182,  428,  1910 

967.  NETTER:  Le  scorbut  infantile.     La  semaine  med.  22  fevr.  No.  8,  1899. 

968.  CHEADLE,  W.  B.  AND  POYNTON  F.  J. :  Infantile  scurvy.    System  of  Med. 

Allbutt-Rolleston    5,  898,  1909. 

969.  LOOSER,  E. :  Ueber  die  Knochenveranderungen  beim  Skorbut.     J.  Kin- 

derh. 62,  743,  1905. 

970.  NEUMANN,   H. :  Bemerkungen  zur  Barlowschen  Krankheit.   D.    med. 

W.  28,  628,  647, 1902;  Sauglingsskorbut.    Deutsch.  Klin.  7,  341,  1904. 

971.  HEUBNER,  O.:  D.  med.  W.  29  Vereinsbeil.  109,  110,  117, 1903;  Ueber  die 

Barlowsche  Krankheit.  Berl.  klin.  W.  40,  285,  1903. 

972.  MEYER,  A.:  Barlows  sygdom.  Kj0benhaven  1901. 

973.  BRACHI  AND  CARR:  Infantile  scurvy  in  a  child  fed  on  sterilized  milk. 

Lane.  662,  1911. 

974.  Amer.  Pediatr.  Soc.,  Collective  investigation  on  infantile  scurvy  in 

North  America.    Arch,  of  Pediatr.  15,  481,  1898. 

975.  HESS,  A.  F.  AND  FISH  M. :  Infantile  scurvy.    The  blood,  the  blood  ves- 

sels and  the  diet.    Amer.  J.  Dis.  Childr.  8,  386,  1914. 

976.  SITTLER:  Das  Sauglingsheim  der  med.     Universitatsklin.   zu  Marburg 

wahrend  der  ersten  Jahre  seines  Bestehens  1905-8  Z.  f.  soc.  Med. 
4,  1,  1909. 


448  THE   VITAMINES 

977.  COMBY,  J. :  Le  scorbut  infantile.    Arch,   de  me"d.   d.  enf ants.  20,  337, 

1917;  22,  281,  1919. 

978.  EPSTEIN,   A. :  Ueber  eine  auffallige  Haufung  der  Barlow'schen  Krank- 

heit  in  den  Kriegsjahren  1917-18  J.   f.   Kinderh.     88,  237,  1918. 

979.  BENDIX:  Barlow'sche  Krankheit.   D.   med.   W.   38,242,1912. 

980.  FORDYCE,  A.  D.:  Scurvy  in  infants.  Edinb.  Med.  J.  20,  110,  1918. 

981.  ASCHOFP,  L.  AND  KOCH.  W.:  Zur  Epidemiologie  des  Skorbuts.    Eine 

pathologisch-anatomische  Studie.  Fischer,   Jena,    1919. 

982.  RHEINDORF:  Med.  Klin.  No.  5,  1918. 

983.  WILTSHIRE,    HAROLD:  Hyperkeratosis   of   the   hair  follicles  in  scurvy. 

Lane.     197,  564,  1919. 

984.  v.  NIEDNER:  Hamorrhagische  Exantheme.    Med.    Klin.  14,  333,  1918. 

985.  WASSERMANN,  S. :  Ueber  eine  mit  Schwellung  und  Rotung  der  Beine 

verbundene  Knochenschmerzhaftigkeit  bei  Kriegern.  Wien.  klin. 
W.  68,  968,  1918. 

986.  ZLOCISTI,  TH.  :  Ueber  Skorbut.  Med.  Klin.  No  25,  659, 1916;  Die  ulzero- 

gangrenosen  Stomatitisf ormen  des  Skorbuts,   Ibid.   No.   46,  1200, 
1916. 

987.  RICHTER,  A. :  Ueber  das  Vorkommen  von  Tetanussymptomen  bei  Skor- 

but.   Wien.  med.  W.  338,  1917. 

988.  URIZIO,  L. :  Skorbut  und  eine  nicht  spirochatogene  Ikterusepidemie. 

Wien.  klin.  W.  No  46,  1449,  1917. 

989.  ZAK,  E. :  Wien.  klin.  W.  30,  592,  1917. 

990.  HIFT:  Beobachtungen   iiber  Skorbut   und   Hemeralopie.    Wien.   klin. 

W.  No.  34,  1918. 

991.  O'SHEA,  H.  V.:  Scurvy.  Practitioner.    Oct-Nov.  1918. 

992.  KITAMURA,  S.  Ein  Beitrag  zur  Kenntnis  der  Netzhautveranderungen 

beim  Skorbut.    D.  med.  W.  No.  9,  1910. 

993.  MILLER:  Infantile  scurvy.     Cleveland  Med.   J.  16,  541,  1917. 

994.  FRANCHETTI,  U. :  Riv.  di  clin.  pediatr.  18,  193,  1920 

995.  COMBY,  J. :  Le  scorbut  chez  le  nourisson.    Med.  1,  673,  1920. 

996.  GERSTENBERGER,  H.  J. :  Amer.  J.  Med.  Sci.  155,  253,  1918. 

997.  COZZOLINO,  O.:  Pediatria27,  321,  407,  477,  1919. 

998.  SCHODEL,    J.    AND   NAUMWERK,    C. :  Untersuchungen  iiber  die  Moller- 

Barlow'sche  Krankheit.     Jena  1900. 

999.  HESS,  A.  F. :  Cardio-respiratory  involvement  in  infantile  scurvy.     Proc. 

Soc.  Exp.  Biol.  Med.,  14,  4,  1916,  J.  A.  M.  A.  68,  235,  1917. 

1000.  ERDHEIM,  J. :  Ueber  das  Barlowherz.  Wien.  klin.  W.  No  49,  1293,  1918. 

1001.  ROSIN:  Hamorrhagischen  Diathesen.  Spez.  Path.  Ther.  inn.  Krankh. 

Kraus-Brugsch  8,  911,    1920. 

1002.  LABOR,   M. :  Ueber   das   Leukozytenbild   bei   Skorbut  und  Tibialgia. 

Wien.  klin.  W.    29,  1069,  1916. 

1003.  FRANKEL,   E.:  Fortschr.     Geb.   d.   Rontgenstr.  7,  No  5-6,   1904;   10, 

No.  1,  1906;  Erganzungsb  18. 

1004.  BROWN,    ALAN:  Some  new  features  in  the  diagnosis  of  scurvy  with 

brief    reference    to    metabolic    changes.   Arch.   Pediatr.    32,   No. 
10,  1915. 


LITERATURE   TO    THE   TEXT  449 

1005.  LABOR,    M:   Eine    Beobachtung   iiber    das    Blutbild    des   Skorbuts. 

Wien.  klin.  W.,  No.  29,  912,  1916. 

1006.  LEITNER,   PH.  :  Beitrage  zur  Hamatologie  und  Klinik    des  Skorbuts. 

Wien.  klin.  W.,  No.  31,  978,  1917. 

1007.  BRANDT,  HEDWIG:   Das  Blut  im  Skorbut.     Arch.  f.  Kinderh.  67,395, 

1919. 

1008.  WASSERMANN,  S. :  Ueber  hochwertige  Erythrozyten  und  Hamoglobin- 

befunde  bei  Kriegern.     M.  med.  W.,  65,  927,   1918. 

1009.  BENOIT,  A. :  Le  sang  dans  le  scorbut.    Arch  d.  malad.  du  coeur.     12, 

241,  1919. 

1010.  ROLLY   AND   OPPERMANN:    Blutzucker.    Bio.    Z.   48,  472,  1913. 

1011.  SCHUMM,    O. :   Untersuchungen    iiber    den    Zuckergehalt    des   Blutes 

unter  physiologischen  und  pathologischen  Verhaltnissen.  III. 
Mitt.  H.  96,  204,  1915. 

1012.  HESS,  A.  F.  AND  KILLIAN.  J.  A.:  Chemistry  of  blood  in  scurvy.     Proc. 

Soc.  Exp.  Biol.  Med.  16,43, 1918;  Hess:  Chemistry  of  blood  in  scurvy, 
especially  its  Ca-content.  Ass.  Amer.  Phys.  June  16th  1919 ;  J.  A. 
M.  A.  Aug.  23rd  1919. 

1013.  BAUMAN,  Louis  AND  HOWARD,  C.  P.:  Metabolism    of    scurvy    in  an 

adult.    Arch.  Internal  Med.  9,  665,  1912. 

1014.  LABBE,  HAGUINEA  AND  NEPREUX:    Biochimie  du  scorbut.    Bull.  soc. 

meU  d.  hop.  44,  1094,  1920. 

1015.  LUST,   F. :   Stoffwechselversuch     bei    Barlowschen  Krankheit.     Berl. 

klin.  W.  49,  862,  1912;    Miinch.  med.  W.  59,  785,  1912. 

1016.  LUST,  F.  AND  KLOCMAN  L. :  Stoffwechsel  bei  Barlowschen  Krankheit 

J.  f.  Kinderh.  75,  663,  1914. 

1017.  MOLL,    L. :  Stoffwechsel    in    Barlowschen   Krankheit.    Mitt.    d.  Ges. 

f.  inn.  Med.  u.   Kinderh.  Wien  18,  1919. 

1018.  FRANK,  M. :    Stoffwechsel    in    Moller-Barlowschen  Krankheit.     J.  f. 

Kinderh.  91,  21,  1920. 

1019.  FEIGENBAUM,  D. :    Ein  Beitrag  zur  Kenntnis  der  Riickenmarkblutun- 

gen  beim  Skorbut.     Wien.  klin.  W.  No.  46,  1455,  1917. 

1020.  SMITH,  W.  JOHNSON:  Scurvy.    System  of  Medicine.    Allbutt-Rolleston, 

5,  879,  1909. 

1021.  FRA'NKEL,  E.:  Archiv  und  Atlas  der  normalen  u.  pathol.  Anatomie 

in  typischen  Rontgenbildern.     Erganzungsband  17,  35,  1908. 

1022.  HESS,  A.  F. :    Focal  degeneration  of  the  lumbar  cord  in  a  case  of  in- 

fantile scurvy.     J.    Inf.    Dis.  23,  438,  1918. 

1023.  BAHRDT,  H.  AND  EDELSTEIN,  F. :  Organanalyse  bei  Barlowschen  Krank- 

heit.    Z.  f.  Kinderh,  9,  415,  1914. 

1024.  PAGET:  Annual  Med.  Rep.  for  year  1912,  Somaliland. 

1025.  SCHULHOF,  W. :  Ueber  die  postskorbutische  Zustande  und  deren  Be- 

handlung.     Wien.  klin.  W.  No  25-26,  1918. 

1026.  DELILLE,  ARMAND:  Le  jus  d'orange  dans  le  traitement  et  la  prophy- 

laxe  du  scorbut  infantile,  d'apres  les  recherches  rdcentes.  J.  des 
pratic.  No.  15,  225,  1919. 


450  THE   VITAMINES 

1027.  HARDEN,  A.,  ZILVA,  S.  S.,  AND  STILL,  G.  F. :    Infantile  scurvy;  the 

antiscorbutic  factor  of  lemon  juice  in  treatment.     Lane.  Jan.  4th, 
1919. 

1028.  RUECK:  Scurvy.    Med.  Rec.  91,  152,  1917. 

1029.  CAUTLEY,  E. :  Rickets ;  GARBOD,  BATTEN   AND  THURSFIELD  :  Diseases 

of  children.     Arnold,     London.     1913. 

1029a.  STRONGMAN,  B.  I.,  AND  BOWDITCH,  H.  I.:  Impressions  Concerning 
etiology  of  rickets  among  Italians.  Bost.  Med.  Surg.  J.  184,  443, 
1921. 

1030.  DENNETT,  R.  H. :  The  use  of  boiled  milk  in  infant  feeding.     J.  A. 

M.  A.  63,  1991,  1914. 

1031.  SCHMORL.:    Verh.  dental-path.  Ges.  58,  1909. 

1032.  SCHWARTZ,  H. :  Craniotabes  and  beading  of  ribs  as  signs  of  rickets. 

Amer.  J.  Dis.  Childr.  19,  384,  1920. 

1032a.  KASSOWITZ,  M.:  Ueber  Rachitis  bei  Neugeborenen.  J.f.Kinderh. 
76,369,1912. 

1033.  MERY  AND  PARTURIER:  Le  rachitisme  congenital.     Bull.  soc.  de  p4d. 

Paris,  fevr.  18,  1919. 

1034.  WIELAND,  E. :  Klinische   und   anatomische   Untersuchungeri  iiber  so 

genannte   angeborene   und   iiber    friihzeitig    erworbene    Rachitis, 
J.  f.  Kinderh.  70,  539,  1909. 

1035.  WIELAND,  E.:  Ueber  das  physiologische  Osteoid  bei Foten und  Neuge- 

borenen und  deren  Bedeutung  fur  die  histologische  Diagnose  der  s.  g. 
angeborenen  Rachitis.     Deutsch.  med.  W.  35,  1520,  1909. 

1036.  SINCLAIR,   JOHN  F. :  Problem  of  the  premature  infant.     Arch.   Ped. 

37,  139,  1920. 

I036a.  CARR,  W.  L. :  Care  of  premature  babies.  Amer.  Ped.  Soc.  Meeting, 
June  2-4,  1921.. 

1037.  HAMILTON,   B.:  Calcium  metabolism  of  premature   infants.     Amer., 

J.  Dis.  Childr.  20  316,  1920. 

1038.  RETTERER,  E.  AND  FISCH  J. :  C.r.  soc.  biol.  80,  182,  1917. 

1039.  OSTHEIMER,  M. :  Fragilitas  ossium.  J.  A.  M.  A.  63,  1997,  1914. 

1040.  BOOKMAN,  A. :  The  metabolism  in  osteogenesis  imperfecta  with  special 

reference  to  calcium.  Amer.  J.  Dis.  Childr.  7,  436,  1914. 

1041.  MCCLANAHAN,  H.  M.  AND  WiLLARD,  W.  W.  i  Osteogenesis  imperfecta 

congenita.  Amer.  Ped.  Soc.  June  16th,  1919;    Amer.  J.  Dis.  Childr. 
19,  181,  1920. 

1042.  SCHABAD,  J.  A.:  Der  Stoffwechsel  bei  angeborener  Knochenbruchig- 

keit.     (Osteogenesis  imperfecta)  Z.  f.  Kinderh.  11,  230,  1914. 

1043.  BRADE-BIRKS  H.  K. :  Bone  condition  analogous  to  rickets  in  a  child 

of  five  months.     Lane.  March  27,  1920. 

1044.  HESS  AND  UNGER:  The  diet  of  the  negro  mother  in  New  York  City. 

J.  A.  M.  A.  70,  900,  1918. 

1045.  JAPHA,  A.:  Kriegsernahrung  in  Beziehung  zur  Rachitis.     Berl.  klin. 

W.  56,  921,  1919. 

1046.  ENGEL:  Rickets  in  Germany;  a  study  of  effect  of  war  in  children. 

Lane.  I,  188,  1920. 


LITERATURE    TO    THE    TEXT  451 

1047.  WEISS,  T. :  Cited  according  to  J.  A.  M.  A.  74,  262,  1920. 

1048.  ADAMS,    JANE    AND    HAMILTON,    ALICE:  A  visit  to  Germany.     Brit. 

J.  Dis  Childr.     16,  July-Sept.  1919. 

1049.  SCHLEE:  Spatrachitis.    Munch,  med.  W.  66,  1349,  1919. 

1050.  STETTER,  K. :  Spatrachitis.  Deutsch.  med.  W.  46,  520,  1919. 

1050a.  SAUER,  H.:  Spatrachitis.     Deutsch.  Zenti.  f.  Chirurg.  162,  356,  1921. 

1051.  SUTTON,   H. :  Manifestation  of  rickets  in  school  children.     Med.   J. 

of  Austr.  2,  190,  1920. 

1051a.  FRANGENHEIM,  P. :  Die  Krankheiten  des  Knochensystems  in  Kindes- 
alter.  F.  Enke.  1913. 

1052.  MULLER,    W.:    Schlatter' sche   Krankheit.     Beitr.    z.  klin.    Chir.  110, 

389,  1920. 

1052a.  PAUS,  N.:  Schlatter's  disease.  Norsk.  Mag.  vor  Laegeridenskaben 
82,  320,  1921. 

1052b.  PARK,  E.  A.,  AND  ROWLAND,  J.:  Dangers  to  life  of  severe  involve- 
ment of  thorax  in  rickets.  Johns  Hopk.  Hosp.  Bull.  32, 101, 1921 . 

1052c.  BRUSA,  P.:  Rachitic  deformity  of  thorax.  Riv.  clin.  pediatr.  19, 
210,  1921. 

1053.  ENGEL,  S. :  Rachitis    in    Beziehung   zum    Wachstumstillstand.    Med. 

Klin.   16,  383,  1920. 

1054.  KARGER,   C. :  M.  f.  Kinderh.  18,  21, 1920. 

1055.  LERI  AND  BECK:  Les    consequences  du  rachitisme  chez  les  adultes. 

Ann.  de  med.  6,  449,  1920. 

1056.  JUARISTI,  V. :  The  rachitic  penis.  Arch,  espanol.  de.  pediatr.  3, 286, 1919. 
1056a.  NEUMANN,  H. :    Ueber  die  Beziehungen  der  Krankheiten  des  Kindes- 

alters  zu  den  Zahnkrankheiten.  S.  Volkmann,  Leipzig,  no.  172; 
1897.  Heubner,  Otto:  Lehrb.  der  Kinderkrankh.  I,  674,  1903; 
Birkenthal:  Berl.  Verlagsanst.  Zweite  Aufl.  p.  48,  1912;  Fleisch- 
mann,  L.:  Cited  according  to  Biedl.  Innere  Secretion,  2nd 
Edition,  1,98,1913. 

1056b.  LICHTENSTEIN,  ALFRED  :  Ueber  die  Geschmacksempfindung  gesunder 
und  rachitischer  Kinder.  J.  f .  Kinderh.  37,  76,  1894. 

1057.  v.  RECKLINGHAUSEN:  Untersuchungen  iiber   Rachitis    und   Osteoma- 

lacie. 

1058.  SCHMORL.  :  Die   pathologische  Anatomic   der   rachitischen   Knochen- 

erkrankung  mit  besonderer  Berticksichtigung  iher  Histologie 
urid  Pathogenese.  Erg.  d.  inn.  Med.  u.  Kinderh.  4,  403,  1909. 

1059.  DIBBELT:  Die  Bedeutung  der  Kalkstoffwechselstorung  fur  die  Entsteh- 

ung  der  Rachitis.    Munch,  med.  W.  57,  2121,  2186,  1910. 

1060.  MARFAN,  BARDOUIN  AND  FEUILLE  :  Lesions  de  la  moelle  osseuse  dans 

le  rachitisme.     C.  r.  soc.  biol.  66,  862,  1909. 

1061.  HUTINEL  AND  'TIXIER:  Modifications  de  la  moelle  osseuse  des  rachi- 

tiques.     Ibid.  66,  946,  1909. 

1062.  KASSOWITZ:  Ueber   Rachitis.     J.  f.  Kinderh.  69,  251,  1909;  II.    Osteo- 

chondritis  rachitica.     Ibid.  75,  194,  334,  489,  581,  1912. 

1063.  HEUBNER:  Lehrb.  der  Kinderkrankheiten.     Leipzig,    1906. 

1064.  POMMER:   Untersuchungen    iiber   Osteomalacie    und    Rachitis.     1885. 


452  THE    VITAMINES 

1065.  RIBBERT,  HUGO:  Betrag  zur  Rachitis.     Deutsch.  med.  W.  39,  8,  1913. 

1066.  HAGENBACH   AND   BURCKHARDT:  Klinische   Beohachtungen   iiber    die 

Muskulatur    der    rachitischen.     J.  f.    Kinderh.  69,  471. 

1067.  BING:  Ueber   atonische   Zustande  der  kindlichen   Muskulatur.    Med. 

Klin.     No.    11,  1907. 
1067a.  BANU,  G. :  Lesions  rachitiques  des  muscles.  Nourrisson.    7,229,1921. 

1068.  MOHR:  Noorden's  Handb.  der  Path.  d.  Stoffw.  2,  865. 

1069.  DU  CASTEL  R. :  Le  thymus  rachitique.     C.  r.  soc.  biol.  65,  725,   1908. 

1070.  STOLZNER:  Ueber  Behandlung  der  Rachitis  mit  Nebennierensubstanz. 

J.  f.  Kinderh.  51,  73,  199;  53,  515,  672. 

1071.  CATTANEO:  Die  Nebennieren  bei  Rachitis.  V.  Kongr.  d.  italien.  Ges. 

f.  Kinderh.  Rom.  1905. 

1072.  GASSMANN,    TH.  :  Untersuchungen    von    gesunden    und    rachitischen 

Knochen.     H.  70,  161,  1910-11. 

1073.  SCHABAD  :  Zur  Bedeutung  des  Kalkes  in  der  Pathologic  der  Rachi- 

tis. V.     Der  Mineralgehalt  gesunder  und  rachitischer    Knochen. 
Arch.  f.  Kinderh.  52,  47,  1909. 

1074.  SIMONINI,  R:  Soc.  med.  chir.  June  11,  1913. 

1075.  ROST,  WILLIAM  L. :  RICKETS,  N.  Y.  Med.  J.  102,  505,  1915. 

1076.  ASCHENHEIM  AND  KAUMHEiMER:  Ueber  den  Aschegehalt  der  Musku- 

latur bei  Rachitischen.     M.  f.  Kinderh.  10,  435,  1911. 

1077.  ASCHENHEIM,  ERICH:  Rachitis  und  Spasmophilie.   I.   J.   f.    Kinderh. 

79,446. 
1077a.  DENIS,  W.,  AND  TALBOT,F.B.:    Calcium  in  blood  of  children.    Amer. 

J.Dis.Childr.21,29, 1921. 
1077b.  HOWLAND,  J.,  AND  KRAMER,  B.:    Calcium  and  phosphorus  in  the 

serum  in  relation  to  rickets.    Am.  J.  Dis.  Childr.  22,  105,  1921. 

1078.  DIBBELT:  Aetiologie  der  Rachitis  und  der  Kalkstoffwechsel.    Deutsch. 

med.   W.   No.  12,  555,  1913. 

1079.  DENTON,  MINNA  C. :  How  much  food  does  it  take  to  supply  us  with  the 

calcium  we  need.     J.  Home  Econ.  10,  168,  1918. 

1080.  ARON,   H. :  Kalkbedarf  und  Kalkaufnahme  beim  Saugling   und   die 

Bedeutung  des  Kalkes  fur  die  Aetiologie  der    Rachitis.    Bio.   Z. 
12,  28,  1908. 

1081.  SCHABAD:  Der  Kalkgehalt  der  Frauenmilch.    J.   f.  Kinderh.  74,  511, 

1911. 

1082.  ORGLER:  Der  Kalkstoffwechsel   des   gesunden  und   des   rachitischen 

Kindes.     Erg.  d.  inn.  Med.  u.  Kinderh.  8,  142,  1912. 

1083.  CRONHEIM   AND   MULLER,  ERICH:  Stoffwechselversuche   an   gesunden 

und  rachitischen  Kindern  mit  besonderer  Berlicksichtigung  des 
Mineralstoffwechsels.  Bio.  Z.  9,  76,  1908. 

1084.  SCHABAD:  Der    Kalkstoffwechsel    bei    Rachitis.    Arch.    f.    Kinderh. 

53,  380,  1910. 

1085.  SCHABAD:  Zur  Bedeutung  des  Kalkes  in  der  Pathologic  der  Rachitis. 

IV-Mitt.Der  P-stoffwechsel  bei  Rachitis.     Arch.    f.  Kinderh.  54, 
83,  1910. 

1086.  FLAMINI:  Arch,  farmacol.  6,  No.  12. 

1087.  PEISER,  AMALIE:  J.  f.  Kinderh.  81,  437,  1915. 


LITERATURE    TO    THE    TEXT  453 

1088.  SCHWARZ:  Der   Stickstoff-  und    Schwefelstoffwechsel    in    Fallen  von 

rachitischem  Zwergwuchs.     J.  f.  Kinderh.  72,  549,  712,  1910. 

1089.  SCHABAD:  2   Falle   von    s.g.    Spatrachitis.     Der   Mineralstoffwechsel 

in  Vergleich  mit  kindlicher  Rachitis.  Mitt.  a.  d.  Grenzg.  d.  Med. 
u.  Chir.  23,  82,  1911. 

1090.  DIBBELT:  Die    experimentelle    Erforschung    der    Rachitis.    Erg.    d. 

wiss.  Med.  2,64,1910-11;  Neue  experimentelle  Untersuchung  iiber 
die  Pathogenese  der  Rachitis.  Deutsch.  med.  W.  38,  316,  1912. 

1091.  MEYER,  LUDWIG  F. :  Zur  Kenntnis  des  Mineralstoffwechsels  im  Saug- 

lingsalter.     Bio.   Z.  12,  422,  1908. 

1092.  TAD  A:    Die    Sauglingsnahrung    "Buttermilch,"    eine    kohlenhydrat- 

reiche  Magermilch.     M.  f.  Kinderh.  4,  118,  1905. 

1093.  ROTHBERG,   Ueber  den  Einfluss  der  organischen  Komponente  auf  den 

Kalkansatz  kiinstlich  genahrten  Sauglinge.  J.  f.  Kinderh.  66, 
69,  1907. 

1094.  ORGLER:  Ueber  den  Kalkstoffwechsel  bei  Rachitis.    M.  f.  Kinderh. 

10,  373,  1911. 
1094a.  HESS,  A.  F.:  Newer  aspects  of  some  nutritional  disorders.     (Harvey 

Lecture)  J.  A.  M.  A.  76,  693,  1921. 
1094b.  TELFER,  S.  V.:  Influence  of  cod  liver  oil  and  butter  fat  on  retention  of 

calcium  and  phosphorus.    Proc.  Physiol.Soc.,  J.Physiol.54,  cv,  1921. 

1095.  HUTCHINSON,  H.  S. :  Is  fat  starvation  a  causal  factor  in  production 

of  rickets?    Glasgow  Med.  J.  11,  8,  1920. 

1096.  HUTCHINSON,  H.  S. :  Fat  metabolism  in  health  and  disease  with  special 

reference  to  infancy  and  childhood.     Quart.  J.  Med.  13,  277,  1920. 

1097.  HOLT,  L.  E.,  COURTNEY,  A.  M.  AND  FALES,  H.  L. :  Fat  metabolism  in 

infants  and  young  children.  III.  Fat  in  stools  of  children  on  mixed 
diet.  Amer.  J.  Dis.  Childr.  18, 107,  1919. 

1098.  HOLT,  L.  E. :  Calcium  metabolism  in  infants  and  young  children,  and 

relation  of  calcium  to  fat  excretion  in  stools.     Ibid.  19,  97,  201,  1920. 

1099.  DIBBELT:  Die  Pathogenese  der  Rachitis.    Arb.  a.  d.  path.  Inst.  Tu- 

bingen 4,  670,  1908. 

1100.  MASSANECK:  Ueber  Buttermilch.     J.  f.  Kinderh.  60,  756,  1904. 

1101.  HOWLAND  AND  MARRIOTT:    Amer.  J.  Obstetr.  74,  541,  1916. 

1102.  SCHABAD,  J.  A. :  Die  gleichzeitige  Verabreichung  von  Phosphorleber- 

tran  mit  einem  Kalksalze  bei  Rachitis.  J.  f.  Kinderh.  72,  1,  1910; 
Z.  f.  klin.  Med.  49,  435,  1910;  Schabad  and  Sorochowetz,  R.  F.: 
M.  f.  Kinderh.  10,  Orig.  2,  12,  1911. 

1103.  SCHABAD:  Dietetic  Treatment  of  rickets.    Russ.    Vratch.  15, 1067,  1916. 

1104.  BIRK,   W. :  Untersuchungen    iiber  den  einfluss  des  P-lebertrans  auf 

den  Mineralstoffwechsel  gesunder  und  rachitischer  Kinder.  M.  f . 
Kinderh.  9,  450,  1908;  Birk  and  Orgler:  Ibid.  9,  544,  1910. 

1105.  FRANK,  L.  AND  SCHLOSS,  E. :  Zur  Therapie  der  Rachitis.  J.  f.  Kinder. 

79,  539,  1914. 

1106.  SCHLOSS,    ERNST:  Deutsch.    med.    W.  39,  1505,   1913;  J.  f.  Kinderh. 

78,  694,  1914;  79,  40,  1914;  Schloss  and  Frank:  Bio.  Z.  60,  378; 
Schloss:  Erg.  d.  inn.  Med.  u.  Kinderh.  15,  1917;  Die  Pathogenese 
und  Aetiologie  der  Rachitis,  sowie  die  Grundlage  ihrer  Therapie. 

1107.  GROSSER,    PAUL  :  Stoffwechselfragen  bei  Rachitis.    Med.  Klin.  10,  577. 


454  THE    VITAMINES 

1108.  Y  ELARDI,  P.  PEREIDA:  Nutritional  derangement  and  rachitis.     Pro- 

gresos   de  la  clin.  7,  117,  1919;    Medic,    ibera  7,  161,  1919. 

1109.  MEYER,    KURT:  Salzstoffwechsel    bei    Rachitis.     J.    f.    Kinderh.  77, 

28,  1914. 

1110.  TOBLER:  Ueber  Spatrachitis.  Munch,  med.  W.  58,  2357,  1911. 

1111.  RICKLIN:  Calcium-   und   Phosphorstoffwechsel    in   einem    Falle    von 

Rachitis  tarda.     Korr.  Blatt.  Schweiz.  Aerzte.  No.  47,  1917. 

1112.  PHEMISTER,  D.  B. :  Effect  of  phosphorus  on  the  growth  of  normal 

and  diseased  bones.     J.  A.  M.  A.  70,  1737,  1918. 
1112a.  PHEMISTER,  D.  B.,   MILLER,  E.  M.  AND  BONAR,  B.  E.:    Effect  of 

phosphorus  in  rickets.    J.  A.  M.  A.  76,  850, 1921. 
1112b.  SHERMAN,  H.  C.  AND  PAPPENHEIMER,  A.  M.:  Dietetic  production  of 

rickets  and  its  prevention  by  an  inorganic  salt.     Proc.  Soc.  Exp. 

Biol.  Med.  18,  193,  1921.    J.  Exp.  Med.  34,  189,  1921. 
1112c.  PAPPENHEIMER,  A.M.,  McCANN,  A.  F.,  ZUCKER,  TH.  F.  AND  HESS, 

A.  F.:    The  effect  of  various  modifications  of  a  diet  producing 

rickets  in  rats.    Proc.  Soc.  Exp.  Biol.  Med.  18;  267, 1921. 

1113.  LIENAUX    AND    HUYNEN:  Bull.    acad.   roy.  med.  beige.  29,  855,  1919. 

1114.  HESS  AND  UNGER:  Prophylactic  therapy  for  rickets  in  a  negro  com- 

munity.   J.  A.  M.  A.  69,  1583,  1917. 

1115.  FERGUSON,  M.:  Medical  Res.   Committee  Rep.   No  20.  A   study  of 

social  and  economic  factors  in  the  causation  of  rickets.  1918. 

1115a.  MACKAY,  HELEN  M.  M.:  Observations  on  cases  of  rickets  in  an  out- 
patient department.  Brit.  Med.  J.  II,  929, 1920. 

1115b.  ARON,  HANS:  Die  Bedeutung  von  Extraktstoffen  fiir  die  Ernahrung. 
II,  M.  f .  Kinderh.  (Orig.)  15,  561,  1920. 

1115c.  RACZYNSKI,  JAN:  Recherches  experimentales  sur  le  manque  d'action 
du  soleil  comme  cause  du  rachitisme.  C.  r.  de  1'assoc.  internat. 
deped.  ler  Congres.  Paris,  1912.  p.  308. 

1115d.  HULDSCHINSKY,  KURT:  Die  Behandlung  der  Rachitis  durch  Ultra- 
violettbestrahlung.  Dargestellt  an  24  Fallen.  Z.  f.  orth.  Chirurg. 
39,  426;Deutsch.med.W.  No.  26, 1919. 

1115e.  MENGERT,  E.:  D.Med.  W.47,  675,  1921. 

1115f.  ERLACHER,  P.:  Phototherapie  bei  Rachitis.  Wien.  klin.  W.  34,  241, 
1921. 

1115g.  HESS,  A.  F.,  AND  UNGER,  L.  J.:  The  cure  of  infantile  rickets  by  sun- 
light. J.  A.M.  A.  77,  39, 1921. 

1115h.  HESS  AND  UNGER:  Interpretation  of  seasonal  variation  of  rickets. 
Proc.  Soc.  Exp.  Biol.  Med.  18,  298,  1921.;  Amer.  J.  Dis.  Childr.22, 
186,  1921. 

1116.  ROWLAND,  JOHN  AND  PARK,  EDWARD  A.:    Amer.  Fed.  Soc.  May  31- 

June  2nd,  1920;  J.  A.  M.  A.  75,  198,  1920. 
1116a.  DUFOUR,  H.:  Bull.  soc.  me*d.  hopit.  Paris.  36, 1261,  1921. 

1117.  v.  HANSEMANN:  Ueber  den  Einfluss  der  Domestikation  auf  die  Ent- 

stehung  der  Krankheiten.     Berl.  klin.  W.  No.  20-21,  1906. 

1118.  KASSOWITZ:  Die  Pathogenese  der  Rachitis.     1885. 

1119.  FINDLAY,  R. :  Rickets.    Brit.  Med.  J.  II,  13,  1908. 


LITERATURE    TO    THE   TEXT  455 

1120.  PATON,  D.  N.,  FINDLAY,  L.  AND  WATSON,  W. :  Observations  on  the  cause 

of  rickets.     Brit.  Med.  J.  II,  625,  1918. 

1121.  PIERSON,  EDITH  AND  DUTCHER,  R.  ADAMS:  Rhubarb  as  an  antiscor- 

butic.    Science,    51,  70,  1920. 

1122.  DICK,   J.  LAWSON:  Defective  Housing  and  the  Growth  of  Children. 

London,  1919;  R.  Soc.  Med.  Febr.  27,  1920.    Lane.  March  13th,  1920. 

1123.  MANN,  H.  CORRY:  88th  Meeting  Brit.  Med.  Ass.  June  30-July  2nd, 

1920.  Brit.  Med.  J.     July  31st,  157,  1920. 
1123a.  CZERNY,  ADALBERT:  Rachitis.    Spez.  Path.  Therap.  inn.  Krankh. 

Kraus-Brugsch.  IX  a,  317, 1921. 
1123b.  SIEGERT,  F.:  Die  Erblichkeit  der  Rachitis.    J.  f.  Kinderh.  58,  929, 

1903. 

1124.  SCHABAD:  Der  Kalk  in  der  Pathologic  der  Rachitis.  Berl.  klin.  w.  46, 

823,  923,  1909. 
1124a.  HERTER,  C.  A.:  J.  Exp.  Med.  3,  293, 1898. 

1125.  HOLT,  L.  E. :  Diseases  of  Infancy  and  Childhood.  1907. 

1126.  CHEADLE,  W.  B.:  Artificial  Feeding  and  Food  Disorders  of  Infants.  1906. 

1127.  STOLZNER.  W. :  J.  f.  Kinderh.  50,  268,  1899. 

1128.  ESSER:  Munch,  med.  W.  1907. 

1129.  PRITCHARD,   ERIC:  Causation   of   rickets   and    its    treatment.     Ped. 

Sept.  1916;  N.  Y.  Med.  J.  110,  921,  1919;    Brit.  Med.  J.  Nov.    15th, 
1919. 

1130.  MELLANBY,    EDWARD:  Accessory   food    factors    (vitamines)    in    the 

feeding  of  infants.     Lane.   I.  856,  1920. 

1131.  MELLANBY,  E.:  R.  Soc.  Med.  Febr.  27,  1920;    Lane.  March  13th,  1920. 
1131a.  PATON,  D.  NOEL  AND  WATSON,  A. :  Etiology  of  rickets.    Brit.  Med.  J. 

1,594, 1921.     Brit.  J.  Exp.  Path.  2,  75,  1921. 

1132.  HOLT,  COURTNEY  AND  FALES:  Fat  metabolism  of  infants  and  young 

children.     IV.  Amer.  J.    Dis.  Childr.  18,  157,  1919. 

1133.  v.   GROER,  FRANZ:  Bio.   Z.  97,  311,  1919. 

1134.  McCoLLUM,  SIMMONDS  AND  PARSONS:  The  etiology  of  rickets.  Proc. 

Amer.    Soc.    Biol.    Chem.    J.    Biol.    Chem.    41,  XXXI,  1920. 

1135.  ROBB:  The  influence  of  dry  vs.  fresh  green  plant  tissue  on  calcium 

metabolism.     Science.   52,  510,  1920. 

1135a.  MCCOLLUM,  SIMMONDS,  PARSONS,  SHIPLEY  AND  PARK:  Studies  on 
experimental  rickets.  I.  The  production  of  rachitis  and  similar 
diseases  in  rats  by  deficient  diets.  J.  Biol.  Chem.  45,  333,  1921. 
45,  343,  1921. 

1135b.  HESS,  McCANN,  AND  PAPPENHEIMER:  The  failure  of  rats  to  develop 
rickets  on  a  diet  deficient  in  vitamine  A.  Soc.  Exp.  Biol.  Med. 
May  18,  1921.  Proc.  18,  266,  1921.  J.  Biol.  Chem.  47,  395,  1921. 

1135c.  TURK,  M.:  Munch,  med.  W.45,  521, 1919. 

1135d.  MCCOLLUM.  E.  V.,  SIMMONDS,  NINA,  SHIPLEY,  P.  G.  AND  PARK,  E.  A.; 
Studies  on  experimental  rickets.  III.  Pathological  condition 
bearing  fundamental  resemblances  to  rickets  of  the  human  being 
resulting  from  diets  low  in  phosphorus  and  fat-soluble  A:  the 
phosphate  ion  in  its  prevention.  Johns.  Hopk.  Hosp.  Bull.  32,  160, 
1921.,  VIII,  J.  Biol.  Chem.  47,  507, 1921. 


456  THE    VITAMINES 

1136.  NATHAN,  M:  Presse  med.  28,  577,  1920. 

1137.  OGATA,   M. :  Rachitis  und  Osteomalacie   in   Japan.     Hegar's    Beitr. 

z.  Geburtsch.     18,  Heft  No.  1. 

1138.  LIESEGANG,  R.  E. :  Puerperal  Osteomalacia.  Zentr.  Gynakol.  39,  No.  15. 

1139.  SCIPIADES,  E.:  Osteomalazie.  Z.  f.  Geburth  u.  Gynakol.  81,  156,  1919. 
1139a.  KOLTONSKI,  H.:  Osteomalazie.    M.  f.  Geburtsh.  u..  Gynakol.  52,  253, 

-    1920. 

1140.  JANUSZEWSKA:  Ueber  Osteomalacie  mit  Anhang  iiber  Tetanic.  Wien. 

klin.  ther.  W.  No.  21,  1910. 

1141.  EDIT.:  Brit.  Med.  J.  Nov.  15th,  1919.     Food  in  Europe  p.  640. 

1142.  PARTSCH,  F.:  Deutsch.  med.  W.  45,  1130,  1919. 

1143.  SAUER:  Ein  Fall  von  Osteomalazie  und  Tetanie  in  Folge  von  Unter- 

ernahrung.     Ibid.  46,  45,  1920. 

1144.  HAMEL,  O. :  Ibid.  46,  68,  1920. 

1145.  HEYER:  Osteomalazie  in  Mimchen.  Munch,  med.  W.  67,  98,  1920. 

1146.  KOPCHEN,  A. :  Z.  f .  inn.  Med.  40,  961,  1919. 

1147.  CURSCHMANN,  H. :  Ueber  Osteomalazie  senilis  und  tarda.  Med.  Klin. 

7,  1565,  1911. 

1147a.  CHELMONSKI,  A.:  Les  maladies  alimentaires  des  os.  Presse.  med.  29, 
115,1921. 

1148.  CROFTAN,     ALFRED    C. :  Osteomalacia.     Amer.     Med.  11,  775,  1916. 

1149.  LOOSER,  E. :  Rachitis  und  Osteomalazia.     Corr.  Bl.  Schweitz.  Aerzte 

49,  1065,  1919. 

1150.  ALWENS:  Die    Beziehungen    der    Unterernahrung    zur     Osteoporose 

und  Osteomalazie.     Miiiich.  rned.  W.  66,  1071,  1919. 

1151.  BOHME,  A.:  Pathologische  Veranderungen  im  Skelett    durch  Unter- 

ernahrung bedingt.     Deutsch.  med.  W.  45,  1160,  1919. 

1152.  IMHOF,  A.:  Psychosen  und  Osteomalazie.    Z.  Neurol.  Psych.  14,  137. 

1153.  GOULD,  ERIC  P. :  The  bone  changes  occurring  in  v.  Recklinghausen's 

disease.     Quart.  J.  Med.  11,  221,  1918. 
1153a.  WEISS,  R.  S.:  v.  Recklinghausen's  disease  in  negro.     Curvature  of 

spine  in  v.  Recklinghausen's  disease.    Arch.  Dermatol.  Syphil.  3, 

144,  1921. 
1153b.  COMBY,  J.:  Bull,  de  Tassoc.  med.  d.  hopit.  45,  382, 1921. 

1154.  HANAU,   A.:   Bericht   iiber  die    Knochen    bei    Osteomalazie.     Korr. 

Bl.   Schweitz.  Aerzte.  497,1892;    Fortschr.   d.   Med.   No.   7,  1892. 

1155.  WILD,    C.    R. :  Anatomische   Untersuchungen   uber    das    puerperale 

Osteophyt.     Diss.  Lausanne,  1901. 

1156.  SCOTT,  AGNES  C. :  Calcium  content  of  the  urine  and  blood  with  reference 

to  its  variation  in  the  condition  of  Osteomalacia.     Ind.  J.  Med. 
4,  169,  1916. 

1157.  ZUNZ,     LEO:    Stoffwechselversuche     bei     der     Osteomalazie.     Arch. 

Gynakol.  99,  145. 

1157a.  ELFER,  A.  AND  KAPPEL,  J. :    Z.  exp.  Path.  Therap.  21,  104,  1920. 
1157b.  HUTCHINSON,  H.  S.,  AND  PATEL,  P.  T.:  Etiology  of  Osteomalacia  in 

Bombay.    Glasgow  Med.  J.  95,  241,  1921. 

1158.  LUZZATTI,  F.:  Spasmophilia  in  children.  Policlin.  26,769,  1919. 


LITERATURE    TO    TfiE    TEXT  457 

1159.  STHEEHMAN,  H.  A.   AND  ARNTZENIUS,  A.  K.  W. :   Signs   of  calcium 

deficiency.     Ned.  Tijdsr.  v.  Gen.   1,  March,  27,  1030,  1920. 

1160.  CYBULSKI:  Ueber  den  Ca-stoffwechsel  der  tetaniekranken  Sauglings. 

M.  f.  Kinderh.  5,  No.  8,  1906. 

1161.  PEPPER,  O.  H.  PERRY:  Disorders  of  nutrition  and  metabolism,  Pro- 

gressive Med.  June  1918. 
1161a.  ROHMER,  P.,  AND  VoNDERWEiDT,  P.  I  Traitement  de  spasmophilie. 

Nourrisson  9,  87,  1921. 
1161b.  SCHERER,   C.  A.:  Manifestations  of  the  spasmophilie  diathesis  in 

older  children.    Minn.  State  Med.  Assoc.    Aug.  24-26,  1921.    J. 

A.  M.  A.  77,966,1921. 

1162.  THIEMICH:  Ueber  die  Behandlung  der  Krampfe  im  friihen  Kindes- 

alter.  Deutsch.  med.  W.  No.  12,  537,    1913. 

1163.  PETRONE,   G.   A.   AND  VITALE,  C. :   Infantile    spasmophilia   and  the 

parathyroid  gland.     La  pediatria  20,  16. 
1163a.  PINCHERLE,  M.,  AND  MAGGESi,  B.  i  Riv.  clin.  pediatr.  18,  577, 1920. 

1164.  TAKASU,    K. :  Spasmophile    Dyspepsie    bei    naturlichen    Ernahrung. 

Sauglingskakke.     J.  f.    Kinderh.  80,    No.   5,    1914. 

1165.  KLOSE,    E.:  Altersgrenzen     ftir     Spasmophilie.     Arch.    f.    Kinderh. 

67,  439,  1919. 

11C6.  BLUHDORN,  K:  Schicksal  der  Kinder  mit  Spasmophilie.  J.  f.  Kinderh. 
92,  294,  1920. 

1167.  FAAS,  JAKOB.  Graviditatstetanie.  Diss.  Erlangen  1913. 

1168.  GUTHRIE:  Tetany;  GARROD,  BATTON  AND   THURSFIELD:  Diseases  of 

Children,  p.  679. 

1169.  LUST,  F. :  Die  Pathogenese  der  Tetanic  im  Kindesalter.  Deutsch.  med. 

W.  No.  23,  1087,  1913. 

1169a.  BOSSERT,  O.  AND  GRALKA,  R.:  Myographie  der  Spasmophilie.  J.  1^ 
Kinderh.  94, 145, 1921. 

1170.  v.    MEYSENBUG,    L. :  Spasmophilia    and    vitamines.     Amer.    J.    Dis, 

Childr.,  20,  206,  1920. 

1170a.  JEPPSON,  J.,  AND  AF  KLERCKER,  K.O.:  Phosphate  bei  der  Spasmo- 
philie. J.  f.  Kinderh.  28,  71,  1921. 

1170b.  IBRAHIM,  J.:  D.  med.  Woch.  46,   1359,  1920. 

1171.  BOSSERT,    O. :  Oedem   bei   der   Spasmophilie.     J.    f.  Kinderh  92,  121, 

1920. 

1172.  ROWLAND,    JOHN  AND  MARRIOTT,    W.   McKra:  Observations   on   the 

calcium  content  of  the  blood  in  infantile  tetany  and  the  effect  of 
treatment  with  calcium.  Bull.  Johns  Hopk.  Hosp.  29,  235,  19181, 
Quart.  J.  J.  Med.  11,  289,  1918. 

1172a.  ELLAS,  H.  AND  SPIEGEL,  E.  A.:  Tetanic.  Wien.  Arch.  inn.  Med. 2, 447, 
1921. 

1172b.  TISDALL,  F.  F.,  KRAMER,  B.  AND  HOWLAND,  J. :  The  concentration  of 
sodium  and  potassium  as  compared  with  that  of  calcium  and  mag- 
nesium in  the  serum  of  patients  with  active  infantile  tetany.  Proc. 
Soc.  Exp.  Biol.  Med.  18,  252, 1921. 

1173.  STHEEHMAN   AND  ARNTZENIUS:   Calcium    level   in   the    blood.    Ned* 

Tijdsr.  v.  Gen.  Apr.  3,  1168,  1920. 


458  THE   VITAMINES 

1173a.  v.  MEYSENBUG,  L.:  Studies  in  spasmophilia  II.  Electrical  reactions 
of  older  children.  Amer.  J.  Dis.  Childr.  20,  539,  1920;  III.  Blood 
calcium  and  calcium  therapy  in  older  children  with  Thiemich's 
sign.  Ibid.  21,  150, 1921. 

1174.  FLETCHER,   ALMON:  Some   considerations   in   the   study   of   infantile 

tetany,  with  report  of  a  case.     Arch.  Internal  Med.  16,  383,  1915. 

1175.  LIEFMANN,    ELSE:  Die   Azetonausscheidung   im    Harn   von   gesunden 

und  spasmophilen  jungen  Kindern.    J.   Kinderh.  77,  125,  1914. 

1176.  SHARPE,  JOHN  SMITH:  The   guanidine  content  of  feces  in   idiopathic 

tetany.     Biochem.  J.  14,  46,  1920. 

1177.  HOOBLER,  B.  RAYMOND:  Calcium-magnesium-phosphorus   balance   in 

children  subject  to  convulsive  disorders.     Amer.  Fed.  Soc.  May 
31-June  2,  1920;    J.  A,  M.  A.  75,  199,  1920. 

1177a.  BOLTEN,  G.  C.:  Electrical  responses  with  Fragilitas  ossium.  Ned. 
Tijdsr.  v.  Gen.  1,  952,  1921. 

1178.  BROWN,  ALAN  AND  FLETCHER,  ALMON:   The  etiology  of  tetany.     Metab- 

olic and  clinical  studies.     Amer.   J.   Dis.   Childr.  10,  313,  1915. 

1179.  STHEEHMAN,    H.    A.:   Spasmophilia     in    children    over    three.     Ned. 

Tijds.  v.   Gen.   No.  14,  Apr.  1,  1145,  1916. 

1180.  BROWN,  A.,  MACL.ACHLAN,  J.  AND  SIMPSON,  F.  R. :  Effect  of  intravenous 

injection  of  Ca  in  tetany  and  influence  of  cod-liver  oil  and  phos- 
phorus in  relation  to  calcium  in  blood.     Amer.   J.   Dis.   Childr. 
19,  413,  1920. 
1180a.  HULDSCHINSKY,  K.:  Z.  f.  Kinderk.  26,  207,  1920. 

1181.  CZERNY:  In  Czerny-Keller.     Handbuch  der  Kinderkrankheiten. 
1181a.  BLOCH,  C.  E.:  Diseases  of  infants  due  to  prolonged  feeding  with 

excess  of  carbohydrates.    Brit.  Med.  J.  I,  293, 1921. 

1182.  RIETSCHEL:  Ueber  Mehle  und  Mehlfutterungen  bei  Sauglingen  und 

ihre  Beziehungen   zurn  Stoffwechsel.     Deutsch.   Med.  W.  34,  826, 
1908. 

1183.  NOEGGERATH:  Zwei  Falle  von  Mehlnahrschaden  beim  Saugling.    Berl. 

klin.  W.  44,  1423,  1907. 

1184.  BOGEN:   Ueber  Mehlnahrschaden.    Deutsch.   Med.   W.  35,  326,  1909. 

1185.  GRUNEBERG:  Nahrschaden  im  Kindesalter.  Munch,  med.  W.  55, 140, 1908. 

1186.  LANGSTEIN,  L:  Jahreskurs  f.  arztl.     Fortb.  3,  24,  1913. 

1187.  MEYER,    L.    F. :  Idiopathische   Oedeme   im   Sauglingsalter.     Erg.    d. 

inn.  Med.  u.  Kinderh.     Bd.  17. 

1188.  RIETSCHEL:    Mehlnahrschaden.    Munch,   med.   W.   No.   19-26,  1918. 

1189.  BENJAMIN:  Mehlnahrschaden.     Z.  f.  Kinderh.  10,  216,  1914. 

1190.  KELLER:  Mehlkinder.    Berl.  klin.  W.  No.  36,  1906. 

1191.  STEINITZ  AND  WEIGERT:  Ueber  den  Einfluss   einseitiger    Ernahrung 

mit    Kohlenhydraten    auf    die    chemische    Zusammensetzung    des 
Sauglingskorpers.     Hofm.  Beitr.  6,  206,  1905. 

1192.  SALGE:  Ein  Beitrag  zur  Pathologic  des  Mehlnahrschadens  der  Saug- 

linge.    Munch,  med.  W.  58,  1915,  1911. 

1193.  ABT,  D.    A.:  Injuries   produced  by  starch.     J.    A.    M.  A.  61,  1275. 

1194.  CAUTLEY:  Atrophia.     Diseases  of  Children.  Garrod,  Batten  &  Thurs- 

field.  London,  1913. 


LITERATURE    TO    THE    TEXT  459 

1195.  FRANK,  A.  AND  STOLTE,  K. :  Mehlnahrschaden.  J.  f.  Kinderh.  78, 167, 191. 

1196.  HOHLFELD:  Rohe  Milch  als  Sauglingsnahrung.   J.  f.  Kinderh.   12,  H. 

1,  1905. 

1197.  BENDIX,    B.:  Chronic    digestive    disturbances    of    infancy.-    Jacobi's 

Diseases  of  Children.     Appleton,  New  York,  1910. 

1198.  MORSE,  J.  L.  AND  TALBOT,  F.  B.:  Diseases  of  Nutrition  and  Infant 

Feeding.   II.   Ed.   Macmillan.  1920,   p.  28. 

1198a.  MARRIOTT,  W.  McK.:  Pathology  of  nutrition  in  infancy.    Amer.  J. 
Dis.Childr.20,461,1920. 

1199.  PEHU:  Alimentation  des  enfants  malades.     Paris,  1908. 

1200.  MORO:  Karottensuppe  bei  Ernahrungstorungen  des  Sauglings.     Munch. 

med.  W.  55,  1637,  1908. 

1201.  STARK,  MORRIS:  Some  studies  on  the  deficiency  diseases  of  infancy 

and    childhood;    interrelation,    prophylaxis,     and    management. 
Amer.  Med.  11,  762,  1916. 

1202.  ARON    AND     SAMELSON:    Karottenextrakt     in     Sauglingsernahrung. 

Deutsch.  med.  W.  46,  772,  1920. 
1202a.  MCCLENDON,  J.  F.  AND  SEDGWICK,  J.  P.:    Powdered  spinach  and  sea 

salts  in  diets  of  adults  and  infants.    Proc.  Soc.  Biol.  Chem.;  J. 

Biol.  Chem.  46,  XXVII,  1921. 
1202b.  ARON,  HANS:  Beitrage  zur  Frage  der  Wirkung  und  Verwertung  der 

Mehle  bei  der  Ernahrung  des  Sauglings.     J.  f.  Kinderh.  42,  82,  1920. 

1203.  KOHLBRUGGE:  Ueber    die    Gahrungskrankheiten.     Centr.     f.     Bakt. 

I.  60,  223,  1911. 

1204.  HOLT,  COURTNEY  AND  FALES:  The  effect  of  cod  liver  oil  on  a  case 

of   "Intestinal  infantilism".     Amer.   J.   Dis.  Childr.  14,222,1917. 

1205.  WEILL  AND  MOURIQUAND:  Lyon  med.  No.  6,  125,  1916. 

1206.  DANIELS,  BYFIELD  AND  LOUGHLIN:  The  role  of  the  antineuritic  vita- 

mine  in  the  artificial  feeding  of   infants.     Amer.  J.  Dis.  Childr. 
18,  546,  1919. 

1207.  EDDYW.  H.  AND  ROPER  J.  C.  :The  use  of  pancreatic  vitamine  in  cases 

of  infant  malnutrition.  Proc.  Soc.  Exp.  Biol.  Med.  14,  52, 1916:  Amer. 
of  J.   Dis.  Childr.  14,  189,  1917. 

1208.  FRIEDBERG,    E.    AND   NOEGGERATH   C. :  Fettfreie   Brustmilch.    Arch. 

f.  Kinderh.  68,  195,  1920. 

1209.  SALGE:  Ueber  Sauglingsatrophie  und  Resorption.    Munch,  med.  W. 

54,  41,  1907. 

1210.  BAHRDT,  H.  AND  EDELSTEIN,  F. :    Energie-  und  Stoffwechsel  in  atro- 

phischen  Saugling.     Z.  Kinderh.  12,  15,  1914. 

1211.  UTHEIM,  K.:  A  study  of  blood  and  its  circulation  in  normal  infants 

and  in  infants   suffering  from  chronic  nutritional   disturbances. 
Amer.  J.  Dis.  Childr.  No.  5,  366,  1920. 

1212.  MATTILL,  P.  M., MAYER,  K.  M.  AND  SAUER,  L.  W.:  Dextrose  tolerance 

in  atrophic  infants.     Ibid.  19,  42,  1920. 

1213.  GLADSTONE,   H.   B. :  Fruit  juices  for  atrophic   infants.     Practit.   97, 

472,  1916. 
1213a.  MARFAN,  A.B.:  Nourrisson.9,65,  1921. 


460  THE    VITAMINES 

1214.  CZERNY,  A.:  M.  f.  Kinderh.  4,  1,  1906;  6,  1,  1908;  7,  1,  1909. 

1215.  SCHIPPERS,  J.  C. :  Fat  content  of  blood.  Ned.  Tijdsr.  v.  Gen.  2, 1081, 1920. 
1215a.  MARFAN.  A.  B.:  Pathogenesis  of  athrepsia.     Nourrisson.  9,  193,  1921. 

1216.  GjG0N,   ALFRED:    Krankheiten    mit    gestortem    Stoffwechsel.     Corr. 

Bl.  f.  Schweiz.  Aerzte.  49,  1529,  1919. 

1217.  GRUMME,    D. :  Der    Nahrwert    der    Proteine.     Therapeut.  Mon.  Nos. 

1-3,  1919. 

1218.  HINDHEDE,  M. :    Adequate  protein  minimum  in  dietaries.  Ugeskr.  for 

Laeger,  79,  Nos.  13-14-15,  1917. 

1219.  HINDHEDE,  M. :  The  protein  minima  on  a  bread  diet.     Skand.  Arch. 

Physiol.  31,  259,  1914;  4th  Rep.  on  protein  minima  with  bread  diet. 
Copenhagen,  Jacob  Lund,  1914. 

1220.  HINDHEDE,  M. :  Ernahrungsversuche  mit  Gerstenwassergriitze.    Skand. 

Arch.    Physiol.     35,  294,  1917. 

1221.  HINDHEDE,  M. :  Fett  in  menschlicher  Ernahrung.  Molk.  Ztg.  28,  152, 

1918;  Fettmiriimum.     Skand.     Arch.  Physiol.  39,  78,  1919. 

1222.  BANG,   S. :  Minimum  ration  of  fat.     Ugeskr.  for  Laeger.  80,  105,  1918. 

1222a.  McCoLLUM,  E.  V.,  SIMMONDS,  NINA  AND  PARSONS,  H.  T.:  Supple- 
mentary protein  values  in  foods  I-V.  J.  Biol.  Chem.  47,  111,  139, 
175,  207,  235,  1921. 

1223.  NEUMANN,  R.  O. :  Kriegsnahrung  in  Bonn  im  Winter  1916-7.   Viert. 

Jahrs.  f.  ger.  Med.  57,  1,  1919. 

1224.  DOLLNER:  Hungerodem.    Munch,  med.  W.  No.  20,  1917. 

1225.  KRASZEWSKI:  Arch.  Hyg.  86,  54,  1916. 

1226.  JANSEN,   W.   H. :  Untersuchungen  iiber  die   Stickstoffbilanz   bei  ka- 

lorienarmen  Nahrung.     D.  Arch.  klin.  Med.  124,  1,  1917. 

1227.  SHERMAN,  H.  C. :  The  protein  requirements  of  maintenance  in  man. 

Proc.  Nat.  Acad.  Sci.  6,  38,  1920. 

1228.  TAYLOR,  A.  E. :  Condition  of  diet  and  nutrition  in  the  Internment 

Camp  at  Ruhleben.     London.  Govern.  Publ.  1916. 

1229.  BENOIT,  ALB.  :    C.  r.  soc.  biol.    82,  151,  1919. 

1230.  BOTAZZI,  F. :  Food  requirements.     Rif .  med.  34,  461,  1918. 

1231.  KRUSE  AND  HINTZE.  Munch,  med.  W.  67,  445,  1920. 

1232.  PRAUSSNITZ,  W.;  Minimum  Nahrungsbediirfnis  des  Menschen.     Ibid. 

67,  696,  1920. 

1233.  STILLE:  Ernahrungslehre  und  Kriegsernahrung.  Schweitzer  &  Co.,  1917. 

1234.  FUNK,  LYLE  AND  MCCASKEY.  :  The  nutritive  value  of  yeast,  polished 

rice  and  white  bread,  as  determined  by  experiments  on  man.     J. 
Biol.  Chem.  27,  173,  1916. 

1235.  HAWK,  P.  B.,  SMITH,  C.  A.  AND  HOLDER,  R.  D. :    Baker's  yeast  as  food 

for  man.    Amer.  J.  Physiol.  48,  199,  1919. 

1236.  RUTGERS,     J. :    Haben     vegetabilischen    Eiweisstoffe    den     gleichen 

Nahrwert  fur    den    Menschen    wie  die  animalischen.     Z.  f.  Biol. 
24,  351,  1888.  . 

1237.  BORUTTAU,  H. :  Wie  wird  pflanzliches  Eiweiss  im  Tierkorper  verwer- 

tet?    Bio.  Z.  69,  225,  1915;  82,  196,  1917. 

1238.  BOTAZZI,  F. :  Importance  des  proteines  et  des  graisses  d'origine  animal 

dans  la  nutrition  del'homme.    Bull.  soc.  d'hyg.  alim.  7,   179,   1919. 


LITERATURE    TO    THE    TEXT  461 

1239.  ROSE,  C.,  AND  BERG,  RAGNAR:   Abhangigkeit   der  Eiweissbediirfnisse 

von  dem  Stoffwechsel.    Miinch.    med.    W.  65,  1011,  1918. 

1240.  HART,  E.  B.  AND  STEENBOCK,  H. :    At  what  level  do  the   proteins  of 

milk  become  effective  supplements  to  the  proteins  of  cereal  grain? 
J.  Biol.  Chem.  42,  167,  1920. 

1241.  MAIGNON,  F. :  C.  r.  soc.  biol.  82,  398,  1919. 

1242.  GRAFE,  E. :  D.  Arch.  klin.  Med.  113,  1. 

1243.  EMMETT,  A.  D. :  Polyneuritis  as  influenced  by  the  amount  of  proteins 

and    carbohydrates    present.     Amer.    Chem.    Soc.    Chicago,  Sept. 
6-10,  1920;  Science,  52,  566,  1920. 

1243a.  OSBORNE  AND  MENDEL:  Results  of  feeding  experiments  with  mixtures 
of  foodstuffs  in  unusual  proportions.  Nat.  Acad.  Sci.  Wash. 
Apr.  26,  1921.  Proc.  Soc.  Exp.  Biol.  Med.  18,  136,  1921;  Growth  on 
diets  containing  more  than  90  per  cent  protein.  Ibid.  18,  167,  1921. 

1244.  TACHAU,  PAUL:  Versuche  tiber  einseitige    Ernahrung.  I.    Mitt.    Bio. 

Z.  65,  253,  1914;  II.  67,  338,  1914. 

1245.  HAUPT,  HUGO:  Chem.  Ztg.  43,  134,  142,  1919. 

1246.  MASON,  C.  C. :  German  nutrition  1914-19.     Bull.  Johns  Hopkins  Hos- 

pital, 31,  66,  1920. 

1247.  BORNSTEIN,  K. :  Berl.  klin.  W.     56,  968,  1919. 

1248.  ZUNTZ,  N.  AND  LOWY,  A.:    Einfluss  der  Kriegsnahrung  auf  den  Stoff- 

wechsel.    Berl.  klin.  W.  53,  825,  1916;  Bio.  Z.  90,  244,  1918. 

1249.  ROSENTHAL,  F. :  Deutsch.  med.  W.  45,  570,  1919. 

1250.  HOFFMAN,  H. :  Arch.  Gynakol.  110,  451,  1919. 

1251.  JAHREISS:  Miinch.  med.  W.  66,  1421,  1919. 

1252.  BLOCK:  Untersuchung  der  unterernahrten  deutschen  Kinder.     Miinch. 

med.  W.  67,  1062,  1920. 
1252a.  KAUPE,   W. :  Muttermilch  und  Kreig.     M.   f.  Kinderh.  15,  83,  1918. 

1253.  ABDERHALDEN,  E. :  Miinch.  med.  W.  Okt.  3,  1919. 

1254.  PFAUNDLER,  M. :  Ueber  Korpermasse  von  Miinchener    Schulkindern 

wahrend  des  Krieges.     Miinch.  med.  W.  Aug.  1,  859,  1919. 

1255.  BLANTON,  S. :  Mental  and  nervous  changes  in  the  school  children  of 

Trier,  Germany,  caused  by  malnutrition.    Mental.  Hyg.  3,  343,  1919. 

1256.  DEMOOR:  Bull.  acad.  roy.  me"d.  beige.  29,  37,   1919;    Demoor  J.  and 

Slosse,  A. :  Ibid.  30,  457,  1920. 
1256a.  DUTHOIT,  RAOUL:  Ibid.  30,  141,  1920. 

1257.  NOBECOURT,  P. :    Presse  med.  27,  653,  1919. 

1258.  FRONCZAK,  F.  E. :  Poland  in  World  War  from  medical  aspect.  N.  Y. 

Med.  J.  Ill,  59,  1920. 

1259.  ROSENFELD,  G. :  Z.  f.  inn.  Med.  41,  305,  1920. 

1260.  BERG,    RAGNAR:  Volksernahrung    im    Kriege.     Blatt,    f.    biol.    Med. 

Sept.  1917. 

1261.  THOMS:  Pharm.  Ztg.  64,  295,  1919. 

1262.  OSBORNE  AND  MENDEL:  Growth  upon  diets  poor  in  true  fats.   Proc. 

Soc.  Exp.  Bio.  Med.  Nov.  17,   1920;  J.  Biol.  Chem.  45,  145,  1920; 
45,  277,  1921. 

1262a.  DRUMMOND,  J.  C. :  Nutrition  on  diets  practically  devoid  of  fat. 
Proc.  Physiol.  Soc.  July  10,  1920.,  J.  Physiol.  44,  xxx,  1920. 


462  THE    VITAMINES 

1263.  HINDHEDE,  M. :  Nahrungsmittelverbrauch  danischer  Familien.    Munch. 

med.  W.  No.  28,  948,  1915;  Deutsch.  med.  W.  No.  45,  1236,  1919; 
Lessons  from  war  restrictions  in  food.  Ugeskr.  for  Laeger.  81, 
183,  1919. 

1264.  KUCZYNSKI,  R.  AND  ZUNTZ,  N. :    Unsere  bisherige  und  unsere  kiinf- 

tige  Ernahrung  im  Kriege.     Vieweg.  1915. 

1265.  HINDHEDE  :  The  influence  of  the  rationing  on  the  health.    Report  to 

the  minister  of  the  interior.  Copenhagen.  1920.  The  effect  of 
restriction  during  the  war  on  mortality  in  Copenhagen.  J.  A.  M.  A. 
74,  381,  1920. 

1266.  NEUHAUS:  Behandlung  der  Falle   durch  Kriegsnahrung  entstanden. 

Deutsch.  med.  W.  No.  25,  1917. 

1267.  NILSSON,  A.:    Z.  f.  Gynakol.  44,  876,  1920. 

1268.  GESSNER,  W. :  Der  Krieg  und  die  puerperale  Eclampsie.    Ibid.  44,  570, 

1920. 

1269.  MOMM:  Munch,  med.  W.  67,  783,  1920. 

1270.  KLOTZ:  Z.  f.  Kinderh.  26,  150,  1920. 

1271.  ZERNIK,  F. :   Kriegsernahrung    und     Arzneimittelwirkung.     Deutsch. 

med.  W.   Juli  31,  858,  1919. 

1272.  KIEFFER,  O. :  Einfluss  des  Krieges  auf  Lungentuberkulose.  Z.  f .  Tuberk. 

32,  65,  1920. 

1273.  Report  of  the  Registrar  General,  for  1917.     Brit.  Med.  J.  May  10,  1919. 

1274.  RUMPEL:  Kriegswirkung  auf  die  Ernahrungsverhaltnisse,    Morbiditat 

und  Mortalitat.    Deutsch.  med.  W.  Juni  19,  1919. 

1275.  ELIAS,   H.   AND  SINGER,  R. :  Diabetes  mellitus  und  Kriegsernahrung 

Deutsch.  med.  W.  46,  561,  1920. 

1276.  SEEFELDER,  R. :  Einfluss  des  Krieges  auf  Augenkrankheiten.   Wien. 

klin.  W.  32,  1245,  1919. 

1277.  HARRIS,    SEALE.  :  Food    conditions    and   nutritional    disturbances  in 

Europe  with  some  remarks  on  the  etiology  of  pellagra.  South. 
Med.  J.  12,  294,  1919. 

1278.  BOUCHARD,  Ch. :  Recherches  nouvelles  sur  la  pellagre.     Paris.    1862. 

1279.  ROUSSEL,  THEOPHILE:  Traite  de  la  pellagre  et  des  pseudopellagres. 

Paris  1866. 

1280.  MARIE,  A. :  La  pellagre.     Paris.     Giard  &  Briere.  1908. 

1281.  ROBERTS,   STEWARD  R. :  Pellagra.   London.   Henry  Kimpton,    1912. 

1282.  SCHILLING,  V. :  Pellagra.  Spez.  Path.  Ther.  inn.  Krankh.  Kraus-Brugsch 

II  a,  72,  1915. 

1283.  NILES,  G.  M. :  Pellagra;  an  American  problem.    2nd  ed.  Saunders  Co. 

1917. 

1284.  HARRIS.  H.  F. :  Pellagra:  A  Study  of  its  Etiology,  Pathology  and  Treat- 

ment, Macmillan  1919. 

1285.  WEISS,  E. :  La  pellagra  nel  Tirolo  meridionale  e  1'azione  del  governo 

contra  la  stessa.  Riv.  pell.  13,  90,  1913;  Oesterr-Sanitatsw.  26,  309, 
1914. 

1286.  SAMBON  AND  CHALMERS:    Pellagra  in  British  Islands.     Brit.  Med.  J. 

p.  1093,  1913. 


LITERATURE   TO   THE   TEXT  463 

1287.  Box  AND  MOTT:  Fatal  pellagra  in  two  English  boys,  with  the  result 

of  the  pathological  investigation  in  one  case.  Trans.  Soc.  Trop. 
Med.  Hyg.  6,  No.  5,  1913. 

1288.  Low,  R.   C.   AND  YELLOWLEES,  H. :  Case  of  pellagra.    Edinb.  Med. 

J.  25,  315,.  1920. 

1289.  PINAULT,  L.  G. :  Pellagra.  J.  Can.  M.  A.   6,  228,  1916. 

1290.  ROLPH,   F.   W.:  Ibid.  6,  323,  1916. 

1291.  LAVINDER:  The  prevalence  and  geographical  distribution   of   pella- 

gra in  the  U.  S.   U.  S.  Publ.  Health  Rep.  Dec.  13, 1912;  28, 1555, 1913. 

1292.  PETERSEN:  Pellagra.  J.  A.  M.  A.  69,  2096,  1917. 

1293.  DEEKS,  W.  E:  Pellagra  in  the  Canal  Zone.    Med.  Rec.  81,  566,  1912; 

South.    Med.    J.  9,  123,  1916. 

1294.  TUTTLE,  H.  K. :  Pellagra  in  Chile.     J.  A.  M.  A.  69,  2105,  1917. 

1295.  MCDONALD,  W.  M. :  Pellagra  in  Antigua.  Lane.  188,  127,  1915. 

1296.  NICHOLLS,   L. :  Tropical  pellagra.   J.  Trop.  Med.  Hyg.  15,  241,  1912. 

1297.  DE  KOCK,  P.  J.  AND  BONNE,  C.  i    Pellagra  in   Surinam.    Ned.  Tijdsr. 

v.  Gen.     2,  965,  1920. 

1298.  ROBERTS,   S.  R. :  Types  and  treatment  of  pellagra.     J.  A.  M.  A.  75, 

21,  1920. 

1299.  SILER  AND  GARRISON:  Amer.  J.  Med.  Sci.  146,  42,  238,  1913. 

1300.  GRIMM:  Pellagra:  some  facts  in  its  epidemiology.  J.  A.  M.   A.  90, 

1423,  1913. 

1301.  WELLMAN  AND  SPARKES:  On  "winter  cases"  of  pellagra.    J.  Trop.  Med. 

Hyg.  15,  131,  1912. 

1302.  SNYDER,  J.  R. :  Pellagra  in  children.  Amer.  J.  Dis.  Childr.  4,  172,  1912. 

1303.  WESTON,    W. :  Pellagra    in    early    childhood.     Ibid.  7,  124,  1914. 

1304.  VOEGTLIN,  C.  AND  HARRIES,  R.  H. :    The  occurrence  of  pellagra  in 

nursing  infants  with  observations  on  the  chemical  composition  of 
the  human  milk  from  pellagrous  mothers.  Hyg.  Lab.  Bull.  Wash. 
116,  73,  1920. 

1305.  BYFIELD,   A.   H. :  Polyneuritic   syndrome   resembling  pellagra   acro- 

dynia  seen  in  young  children.   Amer.  J.  Dis.  Childr.  No.  5,  347, 1920. 

1306.  MURPHY,  W.  A. :  Pellagra  in  children.  Arch.  Fed.  34,  254,  1917. 

1307.  GOLDBERGER,   JOSEPH:    The  etiology  of  pellagra.     The  significance 

of  certain  epidemiological  observations  with  respect  thereto.  U.  S. 
Pub.  Health  Rep.  29,  1683,  1914. 

1308.  WOOD,   EDWARD  J. :  The  diagnosis  of  pellagra.  Arch.  Diagn.  10,  139, 

1917. 

1309.  JOHNSON:  Cited  according  to  Roberts:    Pellagra  (1281). 

1310.  GIVENS,  M.  H. :  Chemical  analysis  of  the  stomachal  content  of   100 

pellagrins.     Amer.   J.  Med.   Sci.  155,  221,  1919. 

1311.  SILER,  J.  F. :  Discussion  on  pellagra.  II.  Pan-Amer.  Sci.  Congr.  Wash. 

1915^16;  Proc.  X,  12,  18,  1917. 

1312.  LYNCH:  Pellagra.  South  Med.  J.  10.  286,  1917. 

1313.  GURD:  Pellagra.  J.  Exp.  Med.  13,  98,  1911. 

1314.  GOLDBERGER,  J. :  Pellagra  and  a  method  of  prevention.  J.  A.  M.  A. 

66,  471,  1916. 


464  THE    VITAMINES 

1315.  DEIACO,  Pius:  Ueber  Lokalisation  und  Natur  der  pellagrosen  Haut- 

symptome.     Wien  klin.  W.  20,  967,  1907. 

1316.  MERK,    LUDWIG:  Die    Hauterscheinungen    bei   Pellagra.     Innsbruck. 

p.  24,  Fig.  6,  1909. 

1317.  CROSBY,   C.   E. :  Pellagra  with  erythema  of  scrotum  as   initial  skin 

manifestation.     J.  A.  M.  A.  68,  1403,  1917. 

1318.  FINDLAY,  G.  M.:    Study  of  the  leucocyte  changes  in  pellagra  com- 

pared with  those  occurring  in  beriberi.    J.  Path.  Bact.  23,  480,  1920. 

1319.  SANDY:  Amer.  J.  Insanity.  73,  609,  1917. 

1320.  MILLER,  R.  S.  AND  ISMAIL,  A.  C. :  Pellagra  and  pellagra  psychoses. 

Lane.  Oct.  16,  788,  1920. 

1321.  CHALMERS,  A.   J.:  Trans.   Soc.   Trop.  Med.   Hyg.  6,  163,  1913. 

1322.  BARDIN,  J. :  Note  on  the  differential  blood  counts  in  three   cases  of 

pellagra.  Old.  Dom.  J.  Med.  &  Surg.  17,  July,  1913. 

1323.  NAGAMATSU,  T. :  Pellagra-like  disease.  Hifukwa,  Hitsunyokwa  Zasshi 

17,  49,  1917. 

1324.  LORENZ,   W.   F. :  The  cerebrospinal  fluid  in  pellagra.   Publ.   Health 

Rep.  29,  2360,  1914. 

1325.  LOMBROSO:  La  pellagra.     Torino  1892. 

1326.  BABES  AND  SIGN:  Cited  according  to  Roberts,  Pellagra.  (1281). 

1327.  CALHOUN:  Ophtalm.  Rec.  26,  63,  1917. 

1328.  BEESON,  CHAS.  F. :  The  thyroid  gland  in  pellagra.     J.  A.  M.  A.  63, 

2129,  1914. 

1329.  MODINOS,  P. :  Bull.   soc.  med.   d.  hop.   de  Paris  Apr.  6,  640,  1916. 

1330.  WILSON:  Appendix  to  report  No  2  on  a  pellagra  epidemic  at  Armenian 

refugees  camp.     Port  Said  Publ.  Health  Dep.  Egypt  1916. 

1331.  MORSE,  PLINN  F. :  The  general  pathology  of  pellagra,  with  special 

reference  to  findings  in  the  thyroid  and  adrenals.     J.  Lab.  Clin. 
Med.  1,  217,  1916. 

1332.  KOCH,  MATHILDE  L.  AND  VOEGTLIN,  C. :    Chemical  changes  in  the  cen- 

tral nervous  system  in  pellagra.     Hyg.  Lab.  Bull.  103,  51,  1916. 

1333.  RIDLON,  J.  R. :  Pellagra.     Laboratory  examination  in  connection  with 

the  disease.     U.    S.    Publ.    Health  Serv.,   No.  339,  1916. 

1334.  MURLIN,  JOHN  R. :  The  amino  acid  fractions  and  hippuric  acid  in  the 

urines  of  pellagrins.     Hyg.  Lab.  Bull.  116,  45,  1920. 

1335.  LEWIS,  ROBERT  C. :  The  chemical  composition  of  the  blood  of  pel- 

lagrins.    Ibid.  116,  37,  1920. 

1336.  JOBLING,  J.  W.  AND  MAXWELL,  E.  S. :    Alkali  reserve  in  the  blood  of 

pellagrins.     J.  A.  M.  A.  69,  2026,  1917. 

1337.  SULLIVAN,  M.  X.  ANDSTANTON,  R.  E. :  Alkali  reserve  in  pellagra.    Arch. 

Internal  Med.  26,  41,  1920. 

1338.  SULLIVAN:  Proc.  Soc.  Amer.  Biol.  Chem.;  J.  Biol.  Chem.  41,  LXX,  1920; 

Sullivan   and   K.   K.    Jones.     The   saliva   in   pellagra.     Ibid.   41, 
LXX,  1920;  Publ.  Health  Rep.  May  16,  1919. 

1339.  MYERS,  VICTOR  C.  AND  FINE,  M.  S. :    Metabolism  in  pellagra.  Amer. 

J.  Med.  Sci.  114,  705,  1913. 

1340.  NICOLAIDI:   Untersuchungen  iiber  Ernahrungsbilanz  der  Pellagrosen. 

Riv.  stiinz.  med.  9,  No.  6,  1913. 


LITERATURE    TO    THE    TEXT  465 

1341.  ALBERTONI,    C.  AND  TULLIO,  P.:     L'alimentazione  maidica    nel  sano 

e  nel  pellagroso.     R.  acad.  scienze  Bologna  11  Jan.  1914. 

1342.  HUNTER,  A.,  GIVENS  AND  LEWIS:  Preliminary  observations  of  metab- 

olism in  pellagra.  U.  S.  Publ.  Health  Serv.  Hyg.  Lab.  Bull.  102, 
39,  1916. 

1343.  BOYD,  F.  D. :  Pellagra.     Edinb.  Med.  J.  24,  366,  1920. 

1343a.  SULLIVAN,  M.  X.,  STANTON,  R.  E.   AND  DAWSON,  P.  R.:    Metabo- 
lism in  pellagra:    Study  of  urine.    Arch.  Int.  Med.  27,  387, 1921. 

1344.  VISWALLINGAM,  A.:Etiol.  of  pellagra.  J.  Trop.  Med.  Hyg.  23,46,  1920. 

1345.  SHEPPARD,  S.  W. :  The  etiol.  of  pellagra.    Brit.    Med.  J.    1773,  1912. 

1346.  STANNUS:  Pellagra    in    Nyasaland.     Trans.    Soc.    Trop.    Med.    Hyg. 

5,  112,  1912;  7,  32,  1913;  Ann.  Med.  Rep.  Nyasaland  1913;  Trop. 
Dis.  Bull.  4,  444,  1914. 

1347.  SCHUFFNER    AND    KuENEN:  Die     gesundheitliche    Verhaltnisse    des 

Arbeiterstandes  des  Sebembah-Maatchappy.  Arch.  Schiffs-u. 
Tropenhyg.  16,  277,  1912. 

1348.  NIGHTINGALE  :  Zeism  or  pellagra?    Brit.  Med.  J.  300,  1914. 

1349. -EDWARDS,  C.  R. :  Peripheral  neuritis  in  Jamaica.  J.  Trop.  Dis.  Hyg. 
19,  53,  1916. 

1350.  WEISS:  Oesterr.     Sanitatsw.  24,  497,  1912. 

1351.  ALPAGO-NOVELLO  :  Pellagra    nel    provincia    di    Belluno.     Riv.    pell. 

ital.  13,  67,  1913. 

1352.  WOOD,   EDWARD  J. :  Vitamine  solution  of  the  pellagra  problem.    J. 

A.  M.  A.  66,  1447,  1916;  Soc.  Trop.  Med.  Hyg.  London.  May 
14,  1920;  Lane.  1166,  1920. 

1353.  BLOSSER,  R. :  Sugar-cane  products  as  a  cause  of  pellagra  in  the  South. 

J.  A.  M.  A.     64,  543,  1915. 

1354.  JOBLING,   J.  W.   AND  PETERSEN,  W.  I    The  epidemiology  of  pellagra 

in  Nashville.    J.  Inf.  Dis.  18,  501,  1916;  21, 109,  1917. 
1355  Babes:  Pellagre.    Bull.  sect.  sci.  de  1'acad.  roumaine.  Nov.  28,  1913. 

1356.  CAMURRI:  Einige  Betrachtungen  iiber  die  Pathogenese  und    die    Be- 

kampfung  der  Pellagra.  Zentr.  f.  Bakt.  I.  Abt.  Orig.  53,  438,  1910. 

1357.  DEVOTO,   L. :  Aetiologie  und  Klinik  der  Pell.     Wien.   med.   W.   No. 

1,  1913. 

1358.  WILSON:  Rep.  of  comm.  appointed  by  the  Direct.  Med.  Serv.  Egypt. 

Exp.  Force,  regarding  the  prevalence  of  pellagra  among  Turkish 
prisoners  of  war.  J.  R.  M.  C.  33,  42t),  508;  34,  70,  1918. 

1359.  BOYD,  F.  D.  AND  LELEAN  P.  S. :  Rep.  of  comm.  of  inquiry  regarding 

the  prevalence  of  pellagra  among  Turkish  prisoners  of  war.  Alex. 
Egypt.  Dec.  31,  1918. 

1360.  LELEAN,  P.  S. :  Pellagra.     Soc.  Trop.  Med.  Hyg.  May  14,  1920;  Lane. 

May  29,  1166,  1920. 

1361.  GOLDBERGER,    J.,    WHEELER,    G.    A.    AND   SYDENSTRICKER   E.  I  Study    of 

the  diets  of  nonpellagrous  and  pellagrous  households.  J.  A.  M.  A. 
71,  944,  1918;  U.  S.  Publ.  Health  Rep.  35,  648,  1920;  Arch.  Internal 
Med.  25,  451,  1920. 


466  THE    VITAMINES 

1362.  SYDENSTRICKER,    EDGAR:  The    prevalence    of    pellagra.     Its    possible 

relation  to  the  rise  in  the  cost  of  food.  U.  S.  Publ.  Health  Rep.  No. 
308,  1915. 

1363.  SULLIVAN  AND  JONES:  The  chemical  composition  of  the  Rankin  Farm 

pellagra-producing  experimental  diets.  Hyg.  Lab.  Wash.  Bull. 
120,  117,  1920. 

1364.  SULLIVAN,  M.  X. :  Ibid.  p.  127,  141. 

1365.  GOLDBERGER:  The  pell,  outbreak  in  Egypt.     Lane.  II.  41,  1920. 

1366.  RATER:  Cited  by  Rayer.  N.  Orl.  Med.  Surg.  J.  66,  718,  1914. 

1367.  KLEIMINGER:  Z.  ges.  Neur.  u.  Psych.  16,  H.  5,  1913. 

1368.  LORENZ  W.  F. :  The  treatm.  of  pell.  U.  S.  Publ.  Hea.  Rep.  29, 2357, 1914. 

1369.  ELEBASH,   C.    C. :  Treating  the   gastro-intestinal    tract    in    pellagra. 

South.  Med.  J.  7,  447,  1914. 

1370.  ALLISON,  W.  L. :  Texas  State  J.  Med.  10,  123,  1914. 

1371.  SYLVESTER,  R.  E.:  Pellagra.     South.  Med.  J.  7,  449,  1914. 

1372.  BRAVETTA,  EUGENIO:  Riv.  pell.  ital.  15,  43,  1915. 

1373.  WILLETS:  South.  Med.  J.  8,  1044,  1915. 

1374.  RIDLON:  Pell.  U.  S.  Publ.  Health  Rep.  No.  353,  1916. 

1375.  GOLDBERGER,  WARING,  C.  H.  AND  WILLETS,  D.  G. :    The  treatment  and 

prevention  of  pellagra.  U.  S.  Publ.  Health  Rep.  No.  228,  No. 
307,  1906.  Goldberger.  Ibid.  No.  461,  1918. 

1376.  VOEGTLIN,  NEILL,  M.  H.  AND  HUNTER,  A. :    The  influence  of  vitamines 

on  the  course  of  pellagra.  Hyg.  Lab.  Bull.  No.  116,  1920;  Voegtlin: 
Harvey  Lecture.  Jan.  24,  1920;  Voegtlin:  Recent  work  on  pellagra. 
U.  S.  Publ.  Health  Rep.  No.  597,  1920. 

1377.  SHERMAN:  Proc.   Amer.   Publ.   Health  Ass.;  Amer.   J.   Publ.  Health, 

10,  86,  1920. 

1378.  SILER,  J.  F.,  GARRISON,  P.  E.  ANDMACNEAL,  W.  J. :  Further  studies 

of  the  Thompson-McFadden  Comm.  J.  A.  M.  A.  63,  1090,  1914. 
1378a.  TANNER,  W.  F.  AND  ECHOLS,  G.  L.:    The  occurrence  of  pellagra  in 
patients  apparently  receiving  an  ample  diet.     J.  A.  M.  A.  76,  1337, 
1921. 

1379.  BOYD,  F.  D. :  Qualitative  problems  of  food  supply.     Lane.  197,  934,  1919. 

1380.  ROAF:  Ibid. 

1381.  BIGLAND,    A.    D. :  Pellagra   outbreak   in   Egypt.    I.     Pellagra    among 

Ottoman  prisoners  of  war.     Lane.  I.  947,  1920. 

1382.  ENRIGHT,  J.  I.:   II.  Pellagra  among  German  prisoners  of  war:  food 

factor  in  disease.     Ibid.  998,  1920. 

1383.  ECKLES,  C.  H.,  PALMER,  L.  S.  AND  SWETT,  W.  O. :    Factors  influencing 

the  composition  of  milk.    Miss.  Agr.  Exp.  Sta.  Bull.  163,  33,  1919. 

1384.  VAN  DER  SCHEER:  Aphtae  tropicae.  Handb.  d.  Tropenk.  von  Mense 

Bd.  II,  S.  1. 

1385.  THIN:  Psilosis  (Sprue)  2nd  Ed.  London  1897. 

1386.  BROWN,  W.  C. :  Sprue  and  its  Treatment.     London  1908. 

1387.  BEGG  :  Sprue,  its  Diagnosis  and  Treatment.     London  1912. 

1388.  WOOD,  EDWARD  J. :  The  recognition  of  tropical  sprue  in  the  United 

States.     Amer.  J.  Med.  Sci.  110,  692,  1915;  J.  A.  M.  A.  73,  165,  1919. 


LITERATURE    TO   THE    TEXT  467 

1389.  BOYD,  M.  F. :  Is  sprue  endemic  in  South?    South.  Med.  J.  13,  229,  1920. 

1390.  STURTEVANT,  M. :  Trop.  sprue  in  New  York.    N.  J.  Med.  Soc.  J.  17, 

44,  1920. 

1391.  HIATT,  H.  B.  AND  ALLAN,  W. :  Notes  on  cases  of  sprue  invalided  from 

the  tropics;  will  it  become  endemic  here?    J.  A.  M.  A.  58,  395,  1914. 

1392.  BAHR,  P.  H. :  Res.  on  sprue.    Trans.  Soc.  Trop.  Med.  Hyg.  7, 161, 1914. 

1393.  HALBERKANN,    J. :  Ham   und   Faces  Untersuchung   bei    der    Sprue. 

Arch.  Schiffs.  Tropenhyg.  20,  225. 

1393a.  BROWN,  T.  B.:  The  absence  of  pancreatic  secretion  in  sprue  and  the 
employment  of  pancreatic  extract  in  the  treatment  of  this 
disease.  Amer.  J.  Med.  Sci.  161,  501, 1921 ;  Bull.  John.  Hopk.  Hosp. 
26,  289, 1916. 

1394.  BASSETT-SMITH,  P.  W. :   Sprue  associated  with  tetany.    Lane.  I.  178, 

1919. 

1395.  BARACH,  A.  L.  AND  MURRAY,  H.   A.:  Tetany  in   a  case  of  sprue.    J. 

A.  M.  A.  74,  786,  1920. 

1396.  SCHMITTER,    F. :  Sprue    treated    by    emetine    hydrochloride.    Milit. 

Surg.  34,  330,  1914. 

1397.  MUHLENS,   P:  Behandlung  bedrohlicher  Zustande   bei  Trope  nkrank- 

heiten.    Deutsch.  med.  W.  40,  1249,  1914. 

1398.  SIMON,   SIDNEY  K. :  Sprue.    South.   Med.   Ass.   Meeting  Nov.   15-18, 

Louisville,  Ky.  1920. 

1398a.  CASTELLANI,  A.:  Treatment  of  sprue  by  massive  doses  of  sodium 
bicarbonate.  Brit.  Med.  J.  I,  338, 1921. 

1399.  WEGELE:  Ueber  die  diatetische  Behandlung  gewisser  Formen  chron- 

ischer  Diarrhoen,   speziell  von   "Indian  Sprue."      Med.  Klin.  9, 
866,  1913. 

1400.  Low,  G.  C. :  Arthritis  in  sprue.     J.  Trop.  Med.  Hyg.  17,  1,  1914. 

1401.  CANTLIE:  Some  recent  observations  on  sprue.    Brit.  Med.  J.  II,  1296, 

1913. 

1402.  CONRAN,  P. :  Meat  diet  in  sprue.     Ibid.  II,  206,  1920. 

1403.  CANTLIE:  The  diet  in  sprue.    J.  Trop.  Med.  Hyg.  9,  277,  1906. 
1403a.  BOVAIRD,  DAVID:  A  study  of  tropical  sprue,  or  psilosis.  J.  A.  M.  A. 

77,753,1921. 

1404.  MICHAEL,  CARL:  A  study  of  toxins  and  the  serological  reactions  in 

sprue.    J.  Amer.  Med.  Sci.  154,  171,  1917. 

1405.  ASHFORD:  Cited  according  to  Siler  (1311). 

1406.  STEWART,  CHARLES  E. :  The  probable  identity  of  pellagra  and  sprue. 

Trans.  17  Intern.  Congr.  Med.  London  Sect.  21,  Trop.  Med.  Part 
II,  125,  1913. 
1406a.  HEATON,  T. :   Etiology  of  sprue.     Ind.  J.  Med.  Res.  7,  810,  1920. 

1407.  WERNER,  H. :  Skorbutsymptome  bei  Sprue.    Arch.  Schiffs.  Tropenhyg. 

No.  7,  1914. 

1408.  LEEDE:  Ein  Fall  von  Sprue  durch  Erdbeeren  gebessert.     Z.  f.  Hyg. 

Infektionsk.  75,  578,  1913. 

1409.  STICKER:  Erkaltungskrankheiten   und   Kalteschaden.     Springer.    Ber- 

lin. 1916. 


468  THE    VITAMINES 

1410.  PRINTING:  Epidemics  resulting  from  wars.   Oxford  Univers.  Press,  1916. 

1411.  MALIWA:  Wien.  klin.  W.  30,  1477,  1917. 

1412.  WHEELER:  War  edema.     Brit.  Med.  J.  No.  2,  1902. 

1413.  DIGBY:  The  famine  campaign  in  Southern  India.     1876-77. 

1413a.  McLEOD,  K. :  Epidemic  dropsy  in  Calcutta.     Ind.  Med.  Gaz.   16, 
148,  1881. 

1414.  PATTERSON,  A.  B. :  Starvation  edema.  Med.  Rec.  Nov.  p.  715,  1899. 

1415.  LANDA:  Deficiency  edema.     Gaceta  med.  Mexico.  11,  67,  1917. 

1416.  RUMPEL:  Kriegsodem.     Munch,     med.    W.    No.    30,    1915;   Berl.    klin. 

W.  1916. 

1417.  JiiRGENs:  Berl.  klin.  W.  No.  9,  1916. 

1418.  BONHEIM:  Munch,  med.  W.  No.  24,  1917. 

1419.  LANGE,  F. :  Deutsch.     med.  W.  No  28,  876,  1917. 

1420.  KNACK:  Wien.  klin.  W.  No.  32,  1916. 

1421.  SCHIFF:  Oedemkrankheit.     Ibid.  No.  22,  1917. 

1422.  JAKSCHE:  Ibid.  68,  1030,  1918. 

1423.  VANDERVELDE,  PAUL  AND  CANTINEAU,  GASTON:    Bull.  acad.  roy.  med. 

belg.  29,  129,  1919. 

1424.  BREUER,  MARCEL:  Ibid.  30,  99,  1920. 

1425.  BEYERMAN,   W. :  "Edema  disease"   in  the  Netherlands,    Ned.  Tijdsr. 

v.  Gen.  1,  2265,  1919. 

1426.  STRAUSS  :  Med.  Klin.  No.  39,  1915. 

1427.  GUILLEMIN,  R.  AND  GUYOT,  F. :  Rev.  med.  de  la  Suisse  romande.  39, 

115,  1919. 

1428.  BUDZYNSKI   B.    AND    CHELCHOWSKI,    K.  i  Hunger   swelling  in  Poland. 

J.  Trop.  Med.  Hyg.  19,  June  15,  1916. 

1429.  WELLS,  H.  G. :  War  edema.     J.  A.  M.  A.  71,  954,  1918. 

1430.  TONIN,  ROMANO:   Gazz.  degli   osped.    e  delle  cliniche.  40,  636,   1919. 

1431.  ENRIGHT,   J.   J. :  War  edema  in  Turkish  prisoners  of  war.     Lane.   I, 

314,  1920. 

1432.  MANN,  W.  L.,  HELM,  J.  B.  AND  BROWN,  C.  J. :  An  edema  disease  in  Haiti. 

J.  A.  M.  A.  75,  1416,  1920. 

1433.  VACHER:  La  mortalite  a  Paris  en  1870.     Gaz.  med.  de  Paris  p.  9,  1871. 

1434.  DE  WOLF,  H. :  A  report  of  13  cases  of  edema  apparently  epidemic  in 

character.  Arch.  Ped.  19,  895,  1902. 

1435.  POTTER,  P.  A. :  The  relation  of  proteins  to  edema  in  marantic  children. 

Med.  News  Jan.  9,  1904;  Arch.  Ped.  29,  206,  1912. 

1436.  CHAPIN,  H.  D. :  Cases  of  edema  in  infants.     Arch.  Ped.  31,  5,  1914. 

1437.  WATERMAN,  L. :  Osmosis  and  edema  in  infancy  and  childhood.  Arch: 

Ped.  31,  135,  1914. 

1438.  HUME,  W.  E. :  General  edema  following  gastro-enteritis  in    children. 

Brit.  Med.  J.  II,  478,  1911. 

1439.  ASHBY,  H.  T. :  Practit.  p.  686,  1914. 

1440.  KLOSE,  ERICH.  :  J.  f .  Kinderh.  80,  154,  1914. 

1440a.  WESTON,  WILLIAM.:  Acrodynia.  Arch.  Ped.  37,  513,  1920. 
1440b.  CARTIN,  H.J.:  Acrodynia.  Penn.  Med.  J.  24,  287, 1921. 

1441.  McCAY,  D. :  Epidemic  dropsy.     Scient.  Mem.  Gov.  India.  No.  37,  204, 

1910. 


LITERATURE    TO    THE    TEXT  469 

1442.  MAVER,  MARIA  B.:  Nutritional  edema  and  "war  dropsy."     J.  A.  M.  A. 

74,  934,  1920. 

1443.  SCHITTENHELM,  A.  AND  SCHLECHT  H. :  Die  Oedemkrankheit.    Springei . 

1919.     Z.  exp.  Med.  9,  1,  1919. 

1444.  SCHIFF,    A.:  Zur  Pathologic    der   Oedemkrankheit.     Wien.    med.    W. 

No.  48,  1917. 

1445.  HULSE,  WALTER:  Munch,  med.  W.  No.  28,  1917. 

1446.  RUMPEL  AND  KNACK:  Berl.  klin.  W.  No.  36,  1916.     Deutsch.  med.  W. 

No.  44,  1916. 

1447.  ZONDEK,  HERMANN  :  Berl.  klin.  W.  55,  502,  1918. 

1448.  JANSEN,  W.  H.:  Munch,  med.  W.  No.  1,  1918. 

1449.  JESS:  Deutsch.  med.  W.  No.  22,  681,  1917. 

1450.  KNACK,  A.  V.  AND  NEUMANN,  I. :  Deutsch.  med.  W.  43,  901,  1917. 

1451.  FALTA:  Wien.  klin.  W.  30,  1736,  1917. 

1452.  BURGER,  MAX:  Epidemisches  Oedem  und  Enterokolitis.     Z.   f.   ges. 

exp.  Med.  8,  309,  1919. 

1453.  KRAUS:  Berl.  klin.  W.  56,  3,  1919. 

1454.  HULSE,  W. :  Virch.  Arch.  225,  234,  1918. 

1455.  LIPPMANN:  Z.  f.  arztl.     Forth.  No.  18,  1917. 

1456.  PALTAUF:  Wien.  klin.  W.  No.  46,  1917. 

1457.  JACOBSTHAL:  Sitzungsb.  arztl.  Ver.  Hamburg.  3  Sept.  1917. 

1458.  WOLTMANN:  Wien.  klin.  W.  1916. 

1459.  MAYNARD,  F.  P. :  Preliminary  note  on  increased  intraocular  tension 

met  with  in  cases  of  epidemic  dropsy.    Ind.  Med.  Gaz.  44,  373,  1909. 

1460.  FRANKE,  M.  AND  GOTTESMANN,  A. :  Wien.  klin.  W.  30,  1004,  1917. 

1461.  FEIGL,  JOHANN:    Bio.  Z.  83,  365,  1918. 

1462.  ZAK:  Wien.  klin.  W.  No.  19,  1917. 

1463.  MAASE   AND   ZONDEK:  Deutsch.   med.   W.   No.   44,  45,  46,   1916;  No. 

16,  1917;  Berl.  klin.  W.  No.  36,  1917. 

1464.  REACH:  Wien.   klin.   W.  31,  1249,  1919. 

1465.  V.HOSSLIN,    HEINRICH:  Klinische    Eigenthumlichkeiten    und    Ernah- 

rung  bei  schweren  Inanition.     Arch.  Hyg.  88,  147,  1919. 

1466.  SCHITTENHELM,  A. :  Eiweisstherapie.  Munch,  med.  W.  66,  1408,  1919. 

1467.  ISENSCHMID,   R. :  Behandlung  von  Oedemen.    Schweiz.  med.  W.  50, 

381,  1920. 

1467a.  ARON,  HANS:  Nahrstoffmangel  als  Krankheitsursache.     Berl.  klin.  W. 
No.  33,  773,  1920. 

1468.  PARK,  F.  S. :  War  edema.      J.  A.  M.  A.  70,  1826,  1918. 

1469.  RUBNER,   M. :  Bericht   an   das  Reichsgesundheitsamt.    Dez.  20,  1917. 

1470.  DETERMANN,  H. :  Die  Bedeutung  der  Kriegsnahrung  fiir  Stoffwechsel 

und  Gesundheit.     Z.   f.   Physik.   u.   diatet.   Ther.  23,  147,  1919. 

1471.  EPSTEIN,  ALBERT  A. :  The  nature  and    treatment    of    chronic  paren- 

chymatous  nephritis.  J.  A.  M.  A.  Aug.  11,  444,  1917.  Amer.  J.  Med. 
Sci.  154,  638,  1917. 

1472.  ALLBUTT:  Dropsy.  Brit.  Med.  J.  II,  395,  1918. 

1473.  DENTON,    MINNA  C.   AND    KOHMAN,    EMMA:     Feeding    experiments 

with  raw  and  boiled  carrots.     J.  Biol.  Chem.  36,  249,  1918. 


470  THE   VITAMINES 

1474.  KOHMAN,  EMMA  A. :  The  experimental  production  of  edema  as  related 

to   protein  deficiency.  Proc.  Soc.  Exp.  Biol.   Med.    16,  121,  1919; 
Amer.  J.  Physiol.  51,  378,  1920. 

1475.  BIGLAND,  A.   D. :  Edema  as  symptom  in  so-called    food    deficiency 

diseases.    Lane.  I.  243,  1920. 

1476.  NIXON,    J.    A.:  Famine  dropsy  as  a  food  deficiency  disease.    Bristol 

Med.  Chirurg.  J.  37,  137,  1920. 

1477.  LITTLE:  Boston  Med.  Surg.  J.  158,  253,  1908;  176,  642,  1917. 

1478.  BRUNTZ,  L.  AND  SPILLMANN,  L. :  C.  r.  soc.  biol.  81, 1243, 1918;  82,  8, 1919; 

Progres  me*d.  34,  9,  1919. 

1479.  MERCIER,  R. :  600  cas  de  froidures  des  pieds.     Bull.  acad.  de  me"d. 

Paris.  82,  80,  1919. 

1480.  CHAUVIN,  E. :  Rev.  de  chir.  38,  793,  1919. 

1481.  McCARRisoN,    R. :  Deficiency   disease:   special   reference    to    gastro- 

intestinal disorders.     Brit.  Med.  J.  I,  822,  1920. 

1482.  LANE,    SIR   WILLIAM   ARBUTHNOT:  Chronic    intestinal    stasis.     Brit. 

Med.    J.  II,  795,  1919. 

1483.  STILL,  G. :  Coeliac  disease.     Lane.  II,  163,  1918. 

1484.  WALLIS,  R.  L.,  MACKENZIE:  88th  Ann.  Meet.     Brit.  Med.  Ass.     Cam- 

bridge, June  30- July  2nd  1920;  Brit.  Med.  J.  160,  1920,  July  31st. 

1485.  REYNOLDS,  EDWARD  AND  MACOMBER,  DONALD:    Deficient   diet   as  a 

cause  of  sterility;  a  study  based  upon  feeding  experiments  with 
rats.    J.A.M.  A.  77, 169, 1921. 

1486.  ISHIWARA:  Nature  of  essential  hemeralopia.     Nippon  Gankakai   Zas- 

shi.     June  1912;  Klin.  Monatsh.  Augenh.  15,  569. 

1487.  TRICOIRE,  R. :  He*meralopie  epid&nique.     Paris  me'd.  10,  152,  1920. 
1487a.  SMITH,  H.:  Night-blindness  and  the  malingering  of  night-blindness. 

J.A.M.  A.  77, 1001, 1921. 

1487b.  APPLETON,  V.  B.:  Observations  on  deficiency  diseases  in  Labrador. 
Amer.  J.  Public  Health.  11,  617, 1921. 

1488.  PICK,  L. :   Einfluss  des  Krieges  auf  die  Augenkrank.     Deutsch.  med. 

W.    46,  44,  1920. 

1489.  FEILCHENFELD,  W. :  Augenkrankheiten  im  Kriege.    Ibid.  46,  575,  1920. 

1490.  HEKTOEN,  LUDWIG:  The  formation  of  antibodies  in  rats  fed  on  pure 

vegetable  proteins  (Osborne-Mendel  diet).  J.  Inf.  Dis.  15,  278,  1914. 

1491.  ZILVA,  S.  S. :  The  influence  of  deficient  nutrition  on  the  production  of 

agglutinins,  complement  and  amboceptor.    Biochem.  J.  13,  172,  1919. 

1492.  KLEINSCHMIDT,  H. :  Ernahrung  und  Bildung  von  Antikorpern.      M.  f . 

Kinderh.  12,  423,  1914. 

1493.  THOMAS,    E. :  Beziehung    zwischen    chronischer    Unterernahrung    zu 

den  klinischen  Zeichen  einerverminderten  Immunitat.  Z.  f.  Kinderh. 
No.  4,  1914. 

1494.  VALAGUSSA,  F. :  Protein  therapy  of  infectious  diseases  of  children. 

Policl.    Med.  Sektion,  27,  361,  1920. 

1495.  PEISER:  Fett  in  der  Kinderdiat.     Berl.  klin.  W.  51,  1065,  1914. 


LITERATURE    TO    THE    TEXT  471 

1496.  Morrison,    J.  R.:  Some  of  the  dangers  of  too  greatly  restricted  diet  in 

typhoid.    Kentucky  Med.  J.  12,  No.  22,  1914. 

1497.  BARKER,  L.  F. :  Diet  in  typhoid  fever.  J.  A.  M.  A.  63,  929,  1914. 

1498.  COLEMAN,  W. :  Influence  of  the  high-caloric  diet  on  the  course    of 

typhoid  fever.     J.  A.  M.  A.  69,  329,  1917. 

1499.  WALTON,  ALICE  C. :  The  new  feeding  in  the  treatment  of  typhoid  fever. 

J.  Home  Econ.  9,  14,  1917. 

1500.  CARTER,  H.  S.,  HOWE,  P.  E.  AND  MASON,  H.  H. :  Nutrition  and  Clini- 

cal Diabetes.     Lea  &  Febiger.     Philadelphia  1917. 

1501.  COMBE,  A.:  Comment  se  nourir  en  temp  de  guerre.  Payot,  Paris.  1917. 

1502.  HERDLIKA,  A. :  Bur.  of  Amer.  Ethnol.  Bull.  34. 

1503.  MCCARRISON,  R. :  88th  Ann.  Meet.  Brit.  Med.  Ass.  Cambridge.  Brit. 

Med.  J.  II,  154,  1920. 

1504.  MORI,  M. :  J.  f .  Kinderh.  59,  175,  1904. 

1505.  CZERNY  AND  KELLER:  Ernahrung  des  Kindes.  p.  2,  67,  1906. 

1506.  KNAPP,  P. :  Experimenteller  Beitrag  zur  Ernahrung  von  Ratten  mit 

kiinstlicher  Nahrung  und  zum  Zusammenhang  von  Ernahrungs- 
storungen  mit  Erkrankungen  der  Conjunktiva.  Z.  f.  exp.  Path, 
u.  Ther.  5,  147,  1909. 

1507.  FREISE,  E.,   GOLDSCHMIDT,   M.   AND  FRANK,  A. :  Experimentelle  Bei- 

trage  zur  Aetiologie  der  Keratomalazie.  Vorl.  Mitt.  M.  f .  Kinderh. 
13,  424,  1914-16. 

1508.  GOLDSCHMIDT,    M. :  Experimenteller   Beitrag  zur  Aetiologie  der  Ke- 

ratomalazie.    Arch.  f.  Ophtal.,  90,  354,  1915. 

1509.  NELSON,  V.  E.  AND  LAMB,  ALVEN,  R. :  Further  studies  on  the  effect 

of  a  deficiency  of  fat-soluble  vitamine.  Amer.  Chem.  Soc.  Chi- 
cago Meet.,  Sept.  6,  1920,  Science  52,  566,  1920. 

1510.  GUERRERO,  L.  E.  AND  CONCEPTION,  I. :  Xerophtalmia  in  fowls  fed  on 

polished    rice  and  its    importance.     Philipp.   J.  Sci.  17,  99,  1920. 

1511.  BLOCK,   C.   E. :  Eye  disease  and  other  disturbances  in  infants   from 

deficiency  in  fat  in  the  food.  Ugeskr.  forLaeger  79,  309,  1917;  80, 
815,  868,  1918;  Ringhosp.  Medd.  2,  17,  1918;  3,  57,  1918. 

1511a.  BLOCK,  C.E.:  Clinical  investigation  on  xerophthalmia  and  dystrophy 
in  young  children.  J.  Hyg.  19,  283, 1921. 

1512.  MONRAD:  Ugeskr.  f.  Laeger.  79,  1177,  1917. 

1513.  R0NNE,  H. :  Ibid.  79,  1479,  1917. 

1514.  SZTARK,   C.   H.:    Arch,  de  malad.    d.    enf.   22,23,1919. 

1515.  PARKER,  R.  H. :   Xerophtalmia.  Iowa  State  Med.  J.  10,  71,  1920. 

1516.  MACFIE,  J.  W.  S. :  Xerophtalmia  in  native  of  Gold  Coast.   Ann.  Trop. 

Med.  Paras.  13,  343,  1920. 

1517.  BULLET,   E.   C.:  Note  on  Xerophtalmia  in  rats.  Biochem.  J.  13,  103, 

1919. 

1518.  STEPHENSON,  M  AND  CLARK,  A.  B. :  A  contribution  to  the  study  of 

keratomalacia  among  rats.  Biochem.  J.  14,  502,  1920. 

1518a.  WASON,  I.M.:  Ophthalmia  associated  with  dietary  deficiency  in  fat- 
soluble  vitamin  A.  Study  of  the  pathology.  J.  A.  M.  A.,  76,  908, 
1921. 


472  THE    VITAMINES 

1519.  EMMETT,  A.  D. :  The  fat-soluble  A-vitamine  and  xerophtalmia.    Science 

52,  157,  1920. 

1520.  MENDEL,  L.  B. :  N.  Y.  Path.  Soc.  Nov.  10,  1920. 

1521.  GUIRAL,  R. :  Keratomalacia  and  its  treatment.     Riv.  med.  y.  cirurg. 

Havana.  24,  157,  1919. 

1522.  HAMBURGER,  RICHARD:  Die  Ernahrung  der  deutschen  Kinder  in  der 

Kriegszeit  und  der  Gegenwart.  Z.  f.  Krankenpfl.     H.  5,  Sept.  1919. 

1523.  STOLZNER,   W. :  Diat  in  Tuberkulose.     Munch,  med.  W.  67,  981,  1920. 

1524.  GEOGHEGAN,   JOSEPH:  Tuberculosis  from  a  West  Indian  standpoint. 

Lane.     July  12th,  1919. 

1525.  RICHET,   CHARLES:  Traitement  de  la  tuberculose  experimentale  par 

la  viande  et  le  serum  musculaire  (zomotherapie)  Trav.  du  lab.  de 
C.  Richet.  5,  1902. 

1526.  WOODCOCK,  H.  de  C.  AND  RUSTIN,  A.  G. :  Food  values  in  tuberculosis. 

Lane.  II,  842,  1920. 

1526a.  GARDEY,  F.:  Vitamines  as  important  factor  in  treatment  of  pul- 
monary tuberculosis.    Semana  med.  27,  759,  1920. 

1527.  WEIGERT:  Berl.  klin.  W.  No.  38,  1904. 

1528.  THOMAS,   ERWIN  AND  HORNEMANN,    O. :  Beziehung  zwischen  der  In- 

fektion  und  Ernahrung.  I.  Bio.  Z.  57,  456. 

1529.  RENON:  La  tuberculose  et  les  vitamines.     Bull.   gen.   de  therap.  30 

Juillet,  1914. 

1530.  MUTHU,  C. :  88th  Meet.  Brit.  Med.  Ass.  Brit.  Med.  J.  II,  160,  1920. 

1531.  BUTTON,  A.  S. :  Some  deficiency  diseases  and  leprosy.     Med.   Press 

N.  S.  109,  313,  1920. 

1532.  HUTCHINSON,  SIR  JONATHAN:  On  leprosy  and  fish  eating.  1906. 

1533.  DEYCKE,  GEORG.  :  Die  Lepra.  Kraus-Brugsch.  Path.  Ther.  d.  Stoffw. 

II,  1,  469. 

1534.  UNDERBILL,  F.  P.,  HONEIJ,  J.  A.,  BOGERT,  L.  J.  ANDALDRICH,  M.  L. : 

Calcium  and  magnesium  metabolism  in  leprosy.     J.   Exp.   Med. 
32,  41,  1920. 
1534a.  VOKURKA:  Korrespondenz  aus  Budapest.    J.  A.  M.  A.  77,  215,  1921. 

1535.  REACH,  FELIX:  Studien  iiber  die  Nebenwirkung  der    Nahrungsstoffe 

Sitzungsb.  Wien.  Akad.  122,  Abt.  Ill,  1,  1913. 

1536.  SALANT,  W. :  The  importance  of  diet  as  a  factor  in  the  production  of 

pathological  change.     J.  A.  M.  A.  69,  603,  1917. 

1537.  SALANT,  W.  AND  SWANSON,  A.  M. :  Further  observations  on  the  influence 

of  diet  on  the  toxicity  of  sodium  tartrate.  Proc.  Soc.  Exp.  Biol. 
Med.  14,  100,  1917. 

1538.  PEARCE,  RICHARD  M.,  AUSTIN,  HAROLD,  J.  AND  PEPPER,  O.  H.  PERRY: 

The  relation  of  the  spleen  to  blood  destruction  and  regeneration 
and  hemolytic  jaundice.  XIII.  The  influence  of  diet  upon  the 
anemia  following  splenectomy.  J.  Exp.  Med.  22,  682,  1915. 

1539.  MADSON,   E.:  The  vitamines.  Ugeskr.  f .  Laeger.  80,  613.  1918. 

1539a.  GEILING,  E.  M.  K.  AND  GREEN,  H.  H.:    Studies  in  regeneration  of 
blood.     Proc.  Soc.  Exp.  Biol.  Med.  18,  191,  1921. 


LITERATURE    TO   THE    TEXT  473 

1540.  DAVIS,  N.  C.,  HALL,  C.  C.  AND  WHIFFLE,  G.  H. :  Rapid  construction 

of  liver  cells  protein  on  a  strict  carbohydrate  diet  contrasted  with 
fasting.     Arch.  Internal  Med.  June  23rd,  689,  711,  1919. 

1541.  CAMPBELL,  H. :  Etiology,  prevention  and  nonoperative  treatment  of 

adenoids.     Brit.   J.   Childr.   Dis.  16,  140,   1919. 

1541a.  VAN  DER  BOGERT,  FRANK  :  Diet  as  a  factor  in  the  etiology  of  adenoids. 
Amer.  Med.  Assoc.  Boston  Meeting.    June  2-10,  1921. 

1542.  HAMMER,  U. :  Deutsch.  med.  W.  46,  738,  1920. 

1543.  PECKAM,  FRANK  E. :  Many  orthopedic  deformities  due  to  calcium  de- 

ficiency, as  direct  result  of  sterilized  and  pasteurized  food.     J.  A. 
M.   A.  75,  1317,  1920. 

1544.  MAGNUS-LEVY,  A.:  Diabetes  in  Kriege.   Deutsch.  med.  W.  45,  1379, 

1919. 

1545.  GERHARDT,    D. :  Diabetes   in    Kriegszeiten.    Schweiz.    med.   W.   50, 

141,  1920. 

1546.  MAGNUS-LEVY:  Diabetes   im   Kraus-Brugsch.  Spez.    Path.    Ther.    d. 

inn.  Krankh.  Bd.  I,  1913. 

1547.  BORUTTAU,   H. :  Spezifische   antidiabetische   Substanz.     Bio.    Z.    88, 

420,  1918. 

1548.  ROSE,  CARL  W. :  Alkaloide  der  Driisen  der  inneren  Sekretion.  Berl. 

klin.  W.  51,  1217. 

1549.  JACOBSEN,  AAGE:  Th.  B.  Bio.  Z.  56,  471. 

1550.  v.  MORACZEWSKI,  W. :  Bio.  Z.  71,  268.  1915. 

1551.  McCAY,  D.,  BANERJEE,  S.  C.,  .GHOSTAL,  L.  M.,  DUTTA,  M.  M.   AND 

RAY,  C. :  .Sugar  of  blood  and  sugar  in  urine  in  varying  conditions 
of  health  in  Bengali.     Ind.  J.  Med.  Res.  6,  485,  508,  1919. 
1551a.  WILDER,  R.  M.  AND  BEELER,  C.:    Plasma  chlorides  and  edema  in 
diabetes.    Amer.  J.  Physiol.  55,  287,  1921. 

1552.  Editorial:   An  unsuspected  occasional  menace     in  low  protein  diets. 

J.  A.  M.  A.  75,  1426,  1920. 

1553.  McCARRisoN,  R. :  Occurrence  of  recently  developed  cancer  of  stomach 

in  monkey  fed  on  food  deficient  in  vitamine.     Ind.  J.  Med.  Res. 
7,  342,  1919. 

1554.  HOFFMAN,  FREDERICK  L. :  The  mortality  from  cancer  in  the  Western 

Hemisphere.     Proc.  II.  Pan-Amer.  Sci.  Congr.  Wash.  X,  586,  1917. 

1555.  FUNK,  CASIMIR  :   The  application  of  chemical  methods  to  the  study  of 

cancer.     Ibid.  X,  388,  1917. 

1556.  EWING,  J. :  Pathological  aspects  of  some  problems  of  experimental 

cancer  research.    J.   Cane.   Res.  1,  71,  1916;  Proc.  II.  Pan-Amer. 
Sci.  Congr.     X,  512,  1917. 

1557.  LOEB,    LEO:  General  problems   and   tendencies   in   cancer   research. 

Ibid.  p.  347. 

1558.  CALKINS,  GARY  N. :  Effects  of  cancer  tissue  and  of  normal  epithelium 

on  the  vitality  of  protozoa.  J.  Cane.  Res.  1,  205,  1916. 

1559.  v.  GRAFF,  E. :  Der  Einfluss  der  Schwangerschaft  auf  das  Wachstum 

maliguer  Tumoren.    Wien.  klin.  W.  27,  7,  1914. 

1560.  SLYE,  MAUD:  J.  Cane.  Res.  5,  25,  1920. 


474  THE    VITAM1NES 

1561.  JANSEN:  Z.  f.  Krebsf.  20,  682,  1909. 

1562.  HAALAND:  Cited  according  to  Ehrlich.  Exper.  Path.  u.  Chemotherapie.. 

Leipzig  1909. 

1563.  CRAMER,  W.  AND  PRINGLE,  HAROLD  :  The  influence  of  diet  on  tumor 

growth.     Proc.  R.  Soc.  88  (B),  307,  315,  1910. 

1564.  Rous,  P.:  The  rate  of  tumor  growth  in  underfed  hosts.    Proc.   Soc. 

Exp.  Biol.  Med.  8,  128,  1911. 

1565.  v.  JAWORSKI,  JOSEF:  Wien.  klin.  W.  p.  1646,  1916. 

1566.  SWEET,  CORSON- WHITE    AND    SAXON:  The  relation  of  diets  and  cas- 

tration on  the  transmissible  tumors  of  the  rats  and  mice.  J. Biol. 
Chem.  15,  181,  1915;  21,  309,  1915.  Corson-White  E.  P. :  Penns.  Med. 
J.  22,  348,  1919. 

1567.  HOPKINS,  F.  G. :  Discussion  on  defic.  diseases.    Proc.  R.  Soc.  Med. 

VII,  1,  (Pharm.  Sect.  )  1913. 

1568.  CENTANNI,  EUGENIC:  La  dieta  aviride  per  lo  sviluppo  dei  tumori  speri- 

mentali.  VI.  Tumori  II,  466,  1914. 

1569.  Rous,  P. :  The  importance  of  diet  in  transplanted  and  spontaneous 

mouse  tumors.     J.  Exp.  Med.  20,  433,  1914. 

1570.  DRUMMOND,  J.  C. :    A  comparative  study   of  tumor  and  normal  tissue- 

growth.     Biochem.  J.  11,  325,  1917. 

1571.  BENEDICT,  S.  R.  AND  RAHE,  ALFRED  H. :  Studies  on  the  influence  of 

various  factors  in  nutrition  upon  the  growth  of  experimental  tumor. 
I.  J.  Cane.  Res.  1,  159,  1917. 

1572.  SUGIURA,  K.  AND  BENEDICT,  S.  R. :  Influence  of  certain  diets  on  tumor 

susceptibility  and  growth  in  albino  rats.  J.  Cane.  Res.  5,  373, 1920. 

1573.  VAN   ALSTYNE  AND  BEEBE:  The  effect  of  non-carbohydrate  diet  upon 

the  growth  of  sarcoma  in  rats.  J.  Med.  Res.  29,  219,  1913. 

1574.  LEVIN,  I. :  Proc.  N.  Y.  Pathol.  Soc.  16,  93,  1916. 

1575.  FRANKEL,  SIGMUND  AND  FURER,  EDINA:  Wien.  klin.  W.  29,  483. 

1576.  RONDONI,  P.:  VII.  Riun.  soc.  ital.  patol.  Pisa  1913. 

1577.  MURPHY:  Transplantability  of  malignant  tumors  to  the  embryo    of 

foreign  species.  J.  A.  M.  A.  59,  874,  1912;  J.  Exp.  Med.  17,  482, 
1913. 

1578.  FUNK,   CASIMIR:  The  transplantation  of  tumors    to  foreign   species. 

J.  Exp.  Med.  21,  571,  1915. 

1579.  BULKLEY,  L.  DUNCAN:  On  the  cure  of  cancer.  Med.  Rec.  97,  941, 1920; 

The  medical  treatment  of  cancer.     Davis  Co.   Phila.   1920. 

1580.  COPEMAN,  S.  MONCKTON:  88th  Ann.  Meet.  Brit.  Med.  Ass.  Brit.  Med. 

J.  II,  159,  1920. 

1581.  KUNERT:  Unsere  heutige  falsche  Ernahrung,  Breslau.  1914. 

1582.  DURAND,  J.  I.:  Influence  of  diet  on  the  development  and  health  of 

teeth.    J.  A.  M.  A.  67,  564,  1916. 

1583.  BLACK,   G.   V.   AND  McKAY,   F.   S. :  Mottled  teeth.    Dent.   Cosmos. 

58,  129,  1916. 

1584.  CASTILLA,  C.  R.:  Changes  in  the  teeth  of  children  from  nutritional 

derangement.     Semana  med.     Buenos  Aires.  26,  599,  1919. 

1585.  MILLER,  E.  G.  AND  GIES,  W.  J. :  Further  nutritive  studies  on  denti- 

tion. J.  Allied  Dent.  Soc.  11,  47,  69,  70,  1916. 


LITERATURE    TO    THE    TEXT  475 

1586.  MELLANBY,  MAY:  The  influence  of  diet  on  teeth  formation.   Lane. 

II,  767,  1918. 

1587.  ZILVA,  S.  S.  AND  WELLS,  F.  M. :  Changes  in  the  teeth  of  the  guinea 

pig  by  a  scorbutic  diet.     Proc.  R.  Soc.  90  (B)  505,  1919. 

1588.  HOWE,   PERCY  R. :  Effect  of    scorbutic  diet  upon  the  teeth.     Dent. 

Cosmos.  62,586,  1920;  62,  921,  1920. 
1588a.  ROBB,  E.  F.,  MEDES,  GRACE;  MCCLENDON,  J.  F.,  GRAHAM,  MARGARET, 

AND  MURPHY,  T.  J.:    A  study  of  scurvy  and  its  bearing  on  the 

preservation  of  the  teeth.    J.  Dent.  Res.  3,  38,  1921. 
1588b.  HOWE,  P.  R.:  Food  accessory  factors  in  relation  to  teeth.    J.  Dental 

Res.  3,  7,  1921. 

1589.  BALLANTYNE:  Brit.  Med.  J.  July  26th,  1919. 

1590.  SINCLAIR,  J.  F. :  Influence  of  diet  affecting  second  dentition.     Penn. 

Med.  J.  12,  789,  1919. 

1591.  OSBORNE  AND  MENDEL:  Incidence  of  phosphatic  urinary  calculi  in 

rats  fed  on  experimental  rations.     J.  A.  M.  A.  69,  32,  1917. 

1592.  PADUA,    REGINO    G. :  Cystolithiasis    among   Filipinos   in   association 

with  dietetic  deficiency.     Philipp.  J.  Sci.  14,  481,  1919. 

1593.  KIRSCHN,ER:  Therapeut.  Monatsh.  33,  300,  1919. 

1594.  CLEMM,  W.  N. :  Ibid.  p.  302; 

1595.  DRUMMOND,  J.  C.:  Vitamines  and  certain  aspects  of  the  relation  to 

public  health.    Amer.  J.  Publ.  Health.  11,  593,  1921. 


INDEX  TO  SECTIONAL  DIVISIONS  OF  THE  LITERATURE 

Historical  Part Nos.  1-68 

Vitamine  Requirements  of  Plants Nos.  69-191 

Vitamine  Requirements  of  Animals Nos.  191a-462 

Chemistry,  Physiology  and  Pharmacology  of  the  Vitamines: 

Vitamine  B Nos.  462a-569 

Vitamine  A Nos.  570-619 

Vitamine  C Nos.  620-672 

Vitamine  Content  of  Various  Foodstuffs Nos.  673-837 

Beriberi Nos.  838-913 

Scurvy Nos.  914-1028 

Rickets,  Osteomalacia,  Tetany,  Spasmophilia  and  Carbohydrate 

Dystrophy Nos.  1029-1215a 

Nutrition  in  Man Nos.  1216-1277 

Pellagra Nos.  1278-1383 

Sprue Nos.  1384-1408 

Hunger  Edema Nos.  1409-1476 

Pathological  Conditions  in  Which  the  Lack  of  Vitamines  may  be 

Suspected .  .Nos.  1477-1595 


476 


AUTHOR  INDEX 


Abderhalden,  20, 25,  37, 
53,  54,  80,  134,  189, 
190,  198,  206,  207, 
211,  255,  256,  264,  265, 
266,  348 

Abt,  336 

Acree,  56 

Adams,  34,  384 

Agulhon,  61 

Alb,  396 

Albert,  294 

Albertoni,  362 

Aldrich,  385 

Alincastre,  196 

Allan,  369 

Allbutt,  376 

Allen,  B.,  245 

Allen,  F.  P.,  85,  86,  121 

Allison,  366 

Almy,  85 

Alpago-Novello,  363 

Alsberg,  395 

van  Alstyne,  390 

Alting,  293 

Alwens,  330 

Amand,  52 

Andersen,  141 

Anderson,  257 

Andrews,  257,  291 

Anrep,  209 

Antoine,  138 

Appleman,  68 

Appleton,  380 

Arima,  293 

Arneth,  299  (#954) 

Arnold,  141 

Arntzenius,  332,  333, 
334 

Aron,  114,  117,  122,  229, 
317,  321,  338,  376,  397 

Aschenheim,  317 


Aschoff,  303,  304,  305, 

308,  309 
Ashby,  373 
Asher,  398 
Ashford,  371 
Aston,  142,  147 
Auer,  256,  265,  266,  267 
Aulde,  397 
Austin,  386 
Autran,  297 
Avery,  62 
Ayers,  57 
Azzi,  396 

B 

Babes,  360,  364 
Bachmann,  54,  198 
Bachstrom,  27,  297 
Balz,  278,  283 
Baglioni,  250,  251 
Bahr,  369 
Bahrdt,  311,  339 
Bainbridge,  56 
Ballantyne,  392 
Bang,  342 
Banner  jee,     388 
Banu,  316 
Barach,  370 
Bardin,  359 
Bardouin,  316 
Barger,  174 
Barker,  381 
Barlow,  27,  257,  302 
Barnes,  259,  270, 
Barsickow,  186 
Bass,  92 

Bassett-Smith,  234,  370 
Bauman,  133,  308 
Baumbach,  299  (#954) 
Baumberger,  84 
Beck,  314 
Beebe,  390 
Beeler,  388 

477 


Beeson,  361 
Beger,  135 
Begg,  369 
Belonowsky,  77 
Bendix,  302,  337 
Benedict,  S.R.,  44, 109, 

125,  195,  269,  389,  390 
Benjamin,  336 
Benoit,  299,  308,  342 
Berg,  343,  348 
van  den  Bergh,  223 
Berthenson,  298 
Bertrand,  56 
Beyerman,  372 
Bezzola,  250 
Bickel,  209 
Bidault,  398 
Bierman,  207 
Bierry,  79,  118,  214 
Biester,  263,  265,  271 
Bigelow,  242 
Bigland,  367,  377 
Bing,  316 
Birk,  320 
BirkenthaL  315 

( # 1056a) 
Black,  391 
Blaine,  297 
Bland-Sutton,  136 
Blantori,  348 
Blatt,  299  (#954) 
Blau,  297 

Bloch,  C.  E.,  335,  382 
Bloch,  348 
Bloombergh,  211 
Blosser,  363 
Blount,  242 
Bluhdorn,  333 
Blunt,  397 
Bogdanow,  77 
Bogen,  335 
Bogert,  385 
van  der  Bogert,  386 


478 


AUTHOR  INDEX 


Bohme,  330 

Bohringer  &  Sons,  186 

Bokorny,  52 

Bolten,  334 

Bonar,  320 

Bonheim,  372 

Bonne,  352 

Bonniger,  136 

Bookman,  314 

Borich,  299 

Bornmann,  245 

Bornstein,  347 

Boruttau,  209,  256,  343, 
387,  395 

Bossert,  333 

Botazzi,  342,  343,  398 

Bottomley,  65,  66,  67, 
68,  69,  70,  71 

Bouchard,  351,  365 

Boutwell,  221,  222,  223, 
224, 239,  254,  258, 265, 
268 

Bovaird,  371 

Bowditch,  312 

Box,  351 

Boyd,  362,  365,  367,  369 

Boye,  138 

Boyer,  60 
Brachi;  302 

Braddon,  29,  210;  212, 

279,  281,  282,  363 
Brade-Birks,  314 
Brandt,  308 
Bravetta,  366 
Breaudat,  31 
Breuer,  372 
Brickman,  260  (#773) 
Brill,  52,  172,  196 
Broomel,  248 
Brown,  A.,  307,  334 
Brown,  C.  J.,  372 
Brown,  T.  B.,  369 
Brown,  W.  C.,  369,  371 
Briining,  114,  122 
Bruntz,  378 
Brusa,  314 
Buckner,  89 
Buckuoy,  297 


Budd,  297 
Budzynski,  372 
Buell,  221,  222 
Biihrer,  396 
Bulkley,  390,  391 
Bull,  138,  220 
Bulley,  382 
Bunge,  21,  22,  26 
Bunker,  60 
Burckhardt,  316 
Burge,  208 
Burger,  374,  388 
Burnet,  245 
Burrows,  82 
Byfield,  41,  43,  123,  157, 
158,  338,  353 


Cahn,  136 
Cajori,  269 
Caldwell,  257 
Calhoun,  361 
Calkins,  79,  389 
Campbell,  H.,  386 
Campbell,  L.  H.,  133 
Campbell,  Mabel,  243, 

266,  267 
Camurri,  364 
Cantineau,  372 
Cantlie,  370,  371 
Carr,  257,  302,  313 
Carrel,  81,  82 
Carter,  381 
Cartin,  373 
Caspari,  280 
du  Castel,  316 
Castellani,  277,  369,  370 
Cattaneo,  316 
Cautley;  312,  336 
Centanni,  389 
Cessna,  55 
Chalmers,  277,  351,  359, 

369 
Chamberlain,    32,    207, 

211,  268,  281 
Chambers,  80 
Chantemesse,  279 
Chapin,  373 


Chapman,  220 

Charrin,  78 

Chauvin,  378 

Cheadle,  301,  302,  307, 
324 

Chelchowski,  372 

Chelmonski,  330 

Chevalier,  278 

Chi  Che  Wang,  397 

Chick,  131,  155,  159, 
233,  236,  238,  243,  245, 
246,  256,  257,  258,  259, 
264,  266,  267,  269, 
270,  301,  367,  398 

Chittenden,  25,  67,  140, 
341,  342 

Christensen,  396 

Chun,  283 

Clark,  296,  385 

Clark,  A.  B.,  383 

Clark,  G.  W.,  50 

Clark,  E.,  92,  93,  103, 
206 

dementi,  251 

Clemm,  393 

Clinton,  65 

Cohen,  131,  241,  265, 
266,  267,  271,  272 

Cohendy,  77,  78 

Cole,    S.   W.,   59,   262, 

270,  271 

Cole,W.  C.C.,  133,  135 
Colebrook,  59 
Coleman,  381 
Collatz,  208 
Combe,    381 
Comby,  302,  306,  330, 

397 

Comrie,  300 
Conception,    258,    270, 

382 

Conklin,  260 
Conran,  370 
Cook  297 
Cooper,  33,  98,  167,  187, 

206,  209,  210,  212,  228, 

229,  237,  255,  262,  266, 

271,  281 


AUTHOR   INDEX 


479 


Copeman,  391 
Coppin,  70 
Coppola,  22 
Corin,  278 
Cornalba,  396 
Corson-White,  389 
Courtney,  319,  327,  338 
Coutts,  260,  270 
Coward,  223,  224,  225, 

226,  265,  266,  269,  270, 

272 

Cowgill,  209 
Cox,  294 
Cozzolino,  307 
Cramer,  229,  230,  389 
Croftan,  330 
Cronheim,  228,  317 
Crosby,  357 
Crowell,  105,  280 
Curjel,  246 
Curran,  262,  298 
Currie,  64 

Curschmann,  330,  331 
Cushny,  209 
Cybulski,  332 
Czerny,  324,  335,  339, 

381 

D 

Dahle,  258  (#754a) 

Dalyell,  158,  159,  238, 
301,  398 

Damianowich,  208 

Daniels,  40,  41,  43,  123, 
157,  158,  240,  241,  243, 
259,  265,  266,  267,  272, 
338 

Darling,  296 

David,  60 

Davis,  M.,  38,  39,  43, 
60,  62,  92,  112,  116, 
220,  221,  224,  245,  259, 
264,  265,  270,  272,  344, 
386 

Davis,  W.  A.,  75,  147 

Dawson,  362 

Dechambre,  297  (#924) 


Decks,  352,  357 

Deiaco,  357 

Delage,  79 

Delcourt,  77 

Delf,  226,  239,  240,  243, 
246,  266,  267,  268 

Delille,  311 

Delpech,  295,  297 

Demoor,  348 

Denis,  317 

Dennett,  312,  317 

Denton,  241,  376 

Desgrez,  118 

Determann,  376 

Devloo,  52 

Devoto,  365 

Deycke,  385 

Dezani,  126 

Dibbelt,  316,  317,  319, 
320 

Dick,  323 

Dienert,  60 

Digby,  372 

Disque",  299 

Djenab,  209 

Dollner,  342 

Doryland,  56 

Douglas,  M.,  105 

Douglas,  S.  R.,  59 

Doyle,  283 

Drescher,  20 

van  Driel,  397 

Driscoll,  251 

Drummond,  39,  43,  88, 
89,  112,  121,  123,  174, 
188,  189,  203,  207,  209, 
221,  222,  223,  224,  225, 
226,  265,  266,  269,  270, 
271,  272,  348,  389,  390, 
393 

Dubin,  55, 117, 123, 124, 
125, 187,  195,  199,  200, 
201,  202,  204,  205,  210, 
214,  215,  227,  234,  246, 
248,  264,  338,  344,  346, 
347,  383,  388,  398 
Dubois,  278 


Dufour,  323 
Dufourt,  301 
Durand,  391 
Diirck,  291 
Durlach,  136  (#400) 
Dutcher,  97,  206,  207, 
208,  245,  258,  263, 268, 

270,  271,  272 
Duthoit,  348 
Dutta,  388 
Dutton,  385 
Dyke,  300 

E 

Eberson,  59 
Echols,  366 
Eckles,  257,  258  (#754- 

a),  368 
Eddy,  55,  79,  189,  196, 

271,  338,  397,  398 
Edelstein,  44,  311,  339 
Edie,  32,  186 
Editorial:     Brit.  Med. 

J.298 
Editorial:    J.  A.  M.  A. 

388 
Editorial:    Trop.   Dis. 

Bull.,  300 
Edwards,  363 
Ehrlich,  389,  390,  391 
Eijkman,  21,  30,  31,  32, 

33,  87,  92,   103,  206, 

211,  212,  245,  397 
Eisenhardt,  209 
y  Elardi,  Pereida,  320 
Elebash,  366 
Elfer,  331 
Elias,  215,  334,  350 
Ellis,  30,  257,  292 
Emmett,  41,  44,  85,  86, 

115, 121, 194,  203,  204, 

238,  271,  346,  383,  398 
Engel,  314 
Engstrand,  133,  135 
Enright,  367,  372 
Epstein,  302,  376 
Erdheim,  307 


480 


AUTHOR   INDEX 


Erlacher,  322 
Esser,  324 
Eustis,  92,  172 
Evans,  186 
Evvard,  142 
Ewald,  206 
van  Eweyk,  209 
Ewing,  266,  389 


Faas,  333 

Faber,  234,  272 

Fahrion,  224 

Fales,  319,  327,  338 

Falk,  242 

Falta,  23,  374,  382 

Feig,  299  (#954) 

Feigenbaum,  309 

Feigl,  375 

Feilchenfeld,  380 

Feuille",  316 

Ferguson,  321,  323 

Ferrari,  299 

Ferry,  43,  60,  74,  112, 

136 
Findlay,  109,  140,  278, 

323,  359 
Fine,  251,  362 
Fingerling,  20 
Fingerling,  A.,  141 
Fink,  103 

Finks,  214,  252,  265,  269 
Fisch,  313 
Fischer,  Emil,  19 
Fischer,  L.,  395 
Fish,  302,  308 
Fitch,  398 
Flack,  58,  92 
Flamini,  318 
Flather,  79,  80 
Fleischmann,  F.,  147 
Fleischmann,    L.,    315 

(#1056a) 

Fleming,  59,  296,  385 
Fletcher,  A.,  334 
Fletcher,  W.,  30 
Fodor,  255 


Folin,  181,  197 

Forbes,  147,  397 

Fordyce,  302 

Forster,  21 

Fowler,  235 

Fraga,  279,  280 

Franchetti,  306 

Frangenheim,  314 

Frank,  A.,  337,  382 

Frank,  L.,  320 

Frank,  M.,  308 

Franke,  375 

Frankel,  E.,  307,  309 

Frankel,  S.,  133,  205, 
390 

Fraser,  30,  31,  93,  94, 
95,  172,  196,  245,  247, 
248,  278,  280,  288,  290 

Freedman,  63 

Freise,  125,  233,  261, 
HIS, 382 

Freudenberg,  225,  233 

Friedberg,  338 

Friedberger,  146 

Froehner,  146 

Frolich,  27,  128,  129, 
131, 134, 137, 144, 231, 
232,  233,  235,  241,  259, 
264,  295,  302 

Fronczak,  348 

Fuji,  109 

Fujitani,  103 

Fuller,  142 

Fulmer,  55 

Funk,  32,  33,  36,  39,  40, 
41,  43,  45,  53,  55,  87, 
88,  89,  96,  97,  98,  105, 
109, 112, 113, 116, 117, 
118,  119, 122,  123,  124, 
125,  130,  133,  134, 
135, 150, 167, 169, 170, 
171,  174,  177,  179,  186, 
187,  192,  195,  196,  197, 
199,200,201,202,203, 
204,  205,  206,  210,  211, 
212,  214,  215,  218,  220, 
227,  233,  246,  248,  252, 


256,  264,  270,  324,  342, 
344,  346,  347,  383,  388, 
389,  390,  395,  396,  398 

Fiirer,  390 

Fiirst,  41,  246 

G 

Gaglio,  210 
Gams,  172 
Garcia,  295 
Gardey,  384 
Garrison,  352,  366 
Gassmann,  317 
Gautier,  219 
Geiling,  386 
Geoghegan,  384 
Gerhardt,  387 
Gerstenberger,  260, 270, 

302,  306 
Gessner,  349 
Ghostal,  388 
Gibson,  258,  270 
Gies,  122,  392 
Gigon,  341 
Gingui,  299 
Givens,    129,  234,  239, 

240,  241,  242,  256,  263, 

266,    267,    268,    271, 

355,  362 
Gladstone,  339 
Goldberger,45,353,357, 

365,  366,  368 
Goldfarb,  81 
Golding,  226 
Goldschmidt,  382 
Gomez,  397 
Gomi,  172 
Goodby,  397 
Gordon,  58 
Gottesmann,  375 
Gould,  330 
Gouzien,  296 
Goy,  205 
Grafe,  E.,  26 
Grafe,  V.,  72 
v.  Graff,  389 
Graham,  392 


AUTHOR   INDEX 


481 


Grnlka,  333 

v.  Hansemann,  323 

Green,  H.  H.,  102,  137, 

Hansen,  23 

141,  144,  146,  148,  150, 

Harden,  40,  43,  55,  85, 

151  (#447),  196,  197, 

123,  129,  154,  193,  233, 

210,  211,  212,  386 

234,  235,  263,  270,  311, 

Green,  Helen  S.,  266 

398 

Greig,    246,    265,    288, 

Harlan,  298 

289,  296,  373 

Harney,  91 

Grenet,  297  (#924) 

Harries,  353,  368 

Grijns,  31,  87,  92,  194, 

Harris,  60,  351 

238 

Harris,  H.  F.,  351 

Grimm,  352 

Harris,  S.,  350 

v.  Groer,  327 

Harrow,  398 

Gross,    249,    264,    266, 

Hart,  C.,  153,  154 

267,  268 

Hart,  E.  B.,  89,  91,  142, 

Grosser,  320 

143,  144,  145,  150,  234, 

Grumme,  341 

257,  260,  270,  343 

Gruneberg,  335 

Harvier,  299 

Guareschi,  248 

Haupt,  347  ' 

Guerrero,  382 

Hausermann,  22,  130 

Guidi,  397 

.  Hawk,  343 

Guillemin,  372 

Hayem,  297  (#924) 

Guillemonat,  78 

Heaton,  371 

Guillerd,  60 

Hebrant,  138 

Guiral,  383 

Hedinger,  149 

Gurd,  357 

Heft,  55,  189 

Guthrie,  333 

Hehir,  279,  281,  299 

Guyenot,  77,  84 

Heim,  130 

Guyot,  372 

Heisig,  240 

Hektoen,  380 

H 

Helm,  372 

Haaland,  389 

Henderson,  140 

Haas,  397,  398 

Henriques,  23,  141 

Haberlandt,  65 

Henry,  147 

Hagenbach,  316 

Hepburn,  278,  283,  294 

Haguinea,  308 

Herdlika,  381 

Halberkann,  369 

Herter,  324 

Hall,  22,  386 

Hertz,  299  (#954) 

Halliburton,    272,    397, 

Hess,  42,  129,  131,  132, 

398 

137,  158,  225,  234,  235, 

Halpin,  20,  89,  91 

240,  242,  243,  258,  260, 

Hamburger,  384 

264,  267,  268,  269,  270, 

Hamel,  330,  331 

271,272,297,298,301, 

Hamilton,  313,  314,  384 

302,  303,  307,  308,  309, 

Hammer,  387 

310,  311,  314,  319,  321, 

Hanan,  330 

322,  326,  327,  328,  336, 

Hannemann,299  (#954) 

339,  387 

Heubner,      Otto,      315 

( #  1056a) 
Heubner,       Wolfgang, 

136,  137,  257,  302,  316 
Hewlett,  155 
Heyer,  330 
Hiat,  369 
Hift,  305 
Hill,  J.  R.,  241 
Hill,  L.,  92,  299 
Hindhede,  241, 245, 255, 

264,  267,  341,  342,  343, 

349,350 
Hine,  58 
Hintze,  342 
Hirota,  257,  290 
Hirsch,  297 
Hoare,  141 
Hocson,  33 
Hogan,  43,  251 
Hoffman,  F.  L.,  388 
Hoffmann,  G.  L.,  129 
Hoffmann,  H.,  347 
Hofmeister,    175,    176> 

177 

Hohlfeld,  337 
Hojer,  397 
Holder,  343 
Holm,  207 
Holmes,  265 
Hoist,  27,  128,  129,  131, 

134,137,144,231,232, 

233,  235,  241,  250,  267, 

295,  302 

Holt,  319,  324,  327,  338 
Honeij,  385 
Hoobler,  334 
Hoogenhuyze,  211,  212 
Hopkins,  F.  G.,  24,  25, 

26,  37,  38,   113,   115, 

116,  125,  223, 224, 256, 

324,  326,  389,  395,  397, 

398 
Hopkins,    G.   R.,   298, 

362 

Hoppert,  150 
Hornemann,  384 


482 


AUTHOR   INDEX 


Horschelmann,  299 

v.  Hosslin,  376 

Houlbert,  92 

Howard,  133,  154,  308 

Howe.  P.  E.,  381 

Howe,  P.  R.,  392 

Rowland,  314,  320,  334 

Hughes,  226,  254 

Huldschinsky,  322,  334 

Hiilse,  373,  374,  375 

Hume,  128,  131,  155, 
233,  236,  238,  245,  246, 
257,  258,  259,  264,  267, 
269,  270,  367,  373 

Humphrey,  56 

Hunt,  386 

Hunter,  109,  362,  366 

Huntoon,  57 

Hutchinson,  H.  S.,  319, 
331,  339 

Hutchinson,  Sir  J.,  385 

Hutinel,  316 

Hutyra,  146 

Hussy,  395 

Huynen,  321 

I 

Ibele,  142 
Ibrahim,  333 
Ide,  55 

Igravidez,  279,  281 
Imhoff,  330 
Ingier,  130 
Ingvaldsen,  154 
Iscovesco,  218,  228 
Isenschmid,  376,  381 
Ishiwara,  379 
Ismail,  358 
Issoglio,  172 
Iwata,  115  (#323) 


Jackson,  111,  114,  129 
Jacob,  23 
Jacobs,  396 
Jacobsen,  388 
Jacobsthal,  374 


Jacoby,  397 

Jahreiss,  348 

Jaksche,  372 

v.  Jaksch,  299  (#954) 

Jansen,  389 

Jansen,  B.  C.  P.,  103, 

207,  208,  269 
Jansen,  W.  H.,  342,  373 
Januszewska,  329,  333 
Japha,  314 
v.  Jaworski,  389 
Jeppson,  333 
Jess,  373 
Jida,  283 
Jizuki,  291 
Jobling,  361,  364 
Johns,  214,  252,  265,  269 
Johnson,  55,  189,  355 
Johnston,  95 
Jones,  69,  141,  261,  361, 

365 

Juaristi,  315 
Judson,  125 
Jiirgens,  372 
Juritz,  253 

K 

Kappel,  331 
Karger,  314 
Karr,  133,  134,  137,  204, 

270 
Kassowitz,     313,     316, 

320,  322,  323 
Kastle,  89 

Katakura,  115  (#323) 
Kato,  104,  283 
Kaumheimer,  317 
Kaupe,  348 
Kaupp,  91 
Kawakami,  146 
Keller,  336,  381 
Kempster,  90,  91 
Kennedy,  39,  222 
Kent,  223,  239,  249,  258, 

264 

Kianizin,  78 
Kieffer,349 


Kilbourne,211 
Killian,308 
Kimura,  103.  104 
King.  135 
Kirschner,  393 
Kitamura,  305 
Kleiminger,  366 
Klein,  143 
Kleinschmidt,  380 
Kleissel,  396 
af  Klercker,  333 
Kligler,  62 
Klocman,  225,  308 
Klose,  333,  373 
Klotz,  349 
Knack,  372,    373,    374, 

376 

Knapp,  382 
Koch,  M.,  109,  361 
Koch,  W.,  303,  304,  305, 

308,  309 
de  Kock,  352 
Kohlbrugge,  338 
Kohler,  53,  80 
Kohman,  376,  377 
Kolb,  215 
Koltonski,  329 
Kondo,  172 
Kopchen,  330,  331 
deKorte,  155  (#459) 
Kossel,  19 
Kramer,   27.    297,   317, 

334 

Kraszewski,  342 
Kraus,  374 
Kraut,  175 
Kriwuscha,  255 
Kruse,  342 
Kubota.  220 
Kuczynski,  349 
Kuenen, 31,238,241,363 
Kulz  103 
Kunert,  391 
Kurijama,  135 
Kurono,  52 
Kusama,  32,  153 
Kiister,  78 


AUTHOR   INDEX 


483 


Labbe,  308 

Labor,  307,  308 

Lagane,  291 

Lake,  119,  135,  137,  245, 

254 

Lamb,  135,  382 
La  Mer,  133 
Lamp6,  25,  37,  134,  206, 

211 

Lampitt,  53 
Lanceraux,  298 
Landa,  372 
Lander.  117 
Lane,    Sir    Arbuthnot, 

379 

Lane-Claypon,  256 
Lange,  372 
de  Langen,  293 
Langstein,  44,  336 
Lasegne,  297  (#924) 
Lavinder,  352 
Leavenworth,  188 
Lecoq,  397 
Leede,  371 
van  Leent,  29 
van  Leersum,  245,  398 
Leggate,  279 
Legroux,  A.,297(#924) 
Legroux,  <R.,  61,  62 
Leitner,  308 
Lelean,  365 
Lemaire,  397 
Le>i,  314 

Leslie,  260  (#773) 
Lessing,  153,  154 
Leven,  297  (#924) 
Levin,  290 
Lewi,  234?  338,  398 
Lewis,    133,    134,    197, 

269,  361,  362 
Lichtenstein,  315 
v.  Liebig,  245 
Liefmann,  334 
Lienaux,  321 
Liesegang,  329 
Lind,  27,  297,  298 


Linossier,  65,  397 
Linton,  397 
Lippmann,  374 
Lipschutz,   136   (#400) 
Little,  279,  378 
Lloyd,  57.  58,  59 
Lobmeyer,  300 
Lockeman,  56 
Loeb,  Jacques,  83,  84 
Loeb,  Leo,  389 
Lombroso,  360,  364,  366 
Looser,  302,  330,  331 
Lorenz,  360,  366 
Lotsch,  146 
Loughlin,    40,    41,    43, 

123,  157, 158,  259,  265, 

266,  272,  338 
Lovelace,  279 
Low,  351,  370 
Lowy,  347 
Luce,  292 
Luckett,  127 
Lucksch,  250 
Lumiere,  79,  102,  208 
Lund,  79 

Lunin,  21,  22,  105 
Luros,  41,  44,  115,  194, 

203,  238,  271 
Lust,  308,  333 
Lutz  64 
Luzzatti,  332 
Lyle,  342 
Lynch,  355 

M 

Maase,  375 

Macalister,  70 

Macallum,  39,  40,  43, 
88,  111,  112,  113,  117, 
118,  119,  123,  181,  197, 
203  270,  344,  383 

Macaulay,  296,  385 

Macauley,  146 

Macfie.  383 

Mackay,  321  m 

Macomber,  379 

MacArthur,  127 


MacDonald,  55 
MacLachlan,  334 
MacNab,  97 
MacNeal  366 
MacPherson,  297 
Madsen,  386 
Magendie,  245 
Maggesi,  333 
Magnus-Levy,  387 
Maignon,   73,   74,   214, 

346 

Maliwa,  372 
Mangkoewinoto,  207 
Mann,  324,  372 
Maranon,  294 
Marek,  146 
Marfan,  316,  339 
Marie,  351,  361,  362 
Marriott,  320,  334,  337 
Martinez,  279 
Mason,  347,  381 
Massalongo,  208,  279 
Massaneck,  320 
Masslow,  137 
Masucci.  60 
Matsunaga,  171 
Matsuyama,  115  (#323) 
Mattei,  248,  270 
Mattill,  122,  261,  339 
Maurer,  210 
Maver,  373 
Maxwell,  361 
Mayer,  339 
Maynard,  375 
McCarrison,    105.    106, 

107,  108,  109,  133,  153, 

207,  216,  217,  227,  377, 

379,  381,  388,  398 
McCann,  321,  327,  328 
McCaskey,  342 
McCay,  373,  388 
McClanahan,  314 
McClendon,    133,    135, 

234,  270,  338,  392,  397 
McClugage,    234,    239, 

240,242,256,263.267, 

268,  271 


484 


AUTHOR   INDEX 


McClurg,  240,  241,  243, 

265,  267 
McCollum,  20,  23,  34, 

38,  39,  40,  43,  44,  55, 
91,  92,  110,  112,  113, 
116,  117, 122, 123, 129, 
130, 131, 134, 142, 143, 
144, 145,  193,  195,  202, 
203,220,221,223,224, 
239,  245,  249,  251, 253, 
255,  257,  259,  264,  265, 

266,  267,  270,  272,  327, 
328,  342,  344,  380,  382, 
396,  397 

McCrae,  253 

McDonald,  352 

McGuire,  242 

McKay,  392 

McKim,  44 

McLean,  228,  268 

McLeod,  372 

Mead,  258  (#754a) 

Medes,  392 

Medical  &  Surgical  His- 
tory of  the  War  of  the 
Rebellion,  297  (#923) 

Meier,  151  (#447) 

Melka,  299  (#954) 

Mellanby,  E.,  42,  138, 
139,  324,  325,  326,  328, 
331,  380 

Mellanby,  M.,  380,  392 

Melocchi,  395 

Meltzer,  340 

Mendel,  37,  38,  39,  40, 
41,  43,  44;  89,  90,  113, 
114,  115,  116,  117,  119, 
121,  122,  123,  125,  127,  . 
131,  195,210,218,221, 
224,227,237,249,251, 
252, 255,  257,  258,  262, 
264,  265,  266,  267,  268, 
269,  270,  271,  272,  34tf. 
348,  382,  383,  393,  395 

Mengert,  322 

Mercier,  378 

v,  Mering,  137 


Merk,  357 
Merklen,  102 
Mery,  313 
Mesnard,  62 
Metchnikoff,  Mme.,  77 
Meyer,  A.;  302 
Meyer,  Kurt,  320 
Meyer,  L.  F.;  319,  336 
v.  Meysenbug,  135,  333, 

334 

Michael,  371 
Michel,  135, 243, 271 
Middlekauff,  133,  135 
Miller,  258,  306 
Miller,  E.  G.,  392 
Miller,  E.  M.,  320 
Miller,  E.  W.,  238,  241 
Miller,  R.;  28 
Miller,  R.  S.,  358 
Miller,  W.S.,  144,  145 
Mitchell,  41,   127,   150, 

203 

Miura,  220,  225,  235, 
278,  283,  284,  285,  286, 
328 

Mockeridge,  70,  71 

Modinos,  361 

Mohr,  316 

Molinari,  398 

Moll,  308 

Momm,  349 

Monrad,  382 

Montagnani,  130,  132 

Moody,  129 

Moore,  129,  186,  258 

v.  Moraczewski,  388 

Morawitz,  299  (#954) 

Morgen,  134 

Morgulis,  85,  122 

Mori,  381 

Morini,  59 

Moro,  77,  338 

Morpurgo   125 

Morrison,  147,  381 

Morse,  301,  337,  361 

Mosc,  396 

Moszkowski,  280 


Mott,  292,  358,  359 

Moufang,  52 

Mourges,  219 

Mouriquand,  78,  92, 
104,  135,  195,  212,  239, 
243,  245,  246,  248,  254, 
264,  271,  272,  291,  299, 
338,  396,  397 

Muckenfuss,  210 

Mueller,  62,  344 

Muhlens,  370 

Muller,  223 

Muller,  298 

Muller,  Erich,  301,  317 

Muller,  W.,  314 

Mulvany,  278 

Munk,  245 

Murai,  172 

Murlin,241,361 

Murray,  370 

Muthu,  385 

Myers,  C.  N.,  96,  97, 
191,  196,  209,  229,  245, 
246,  249,  254 

Myers,  Victor,  362 

Murphy,  T.  J.,  392 

Murphy,  W.  A.,  353 

Murphy,  390 

N 

Nagamatsu,  360 
Nagasawa,  271  ( #836) 
Naish,  260 
Nambu,  297,  303 
Nansen,  262 
Nathan,  328 
Naumwerk,  307,  387 
Neill,  208,  366 
Nelson,  B.  A.,  127 
Nelson,  E.  M.,  222 
Nelson,  V.  E.,  55,  13o, 

257,  382 
Nemoto,  283 
Neppi,  398 
Nepreux,  308 
Netter,  301 
Neuhaus,  349 


AUTHOR   INDEX 


485 


Neumann,  257,  302,  315 

(#1056a),  342,  374 
v.  Neusser,  250 
Neville,  38 
Nichols,  265 
Nicholls,  352 
Nicolaidi,  362 
Nicolet,  197 
v.  Niedner,  303 
Nightingale,  363 
Niles,  351,  353 
Nilsson,  349 
Nitzesco,  251 
Nixon,  377 
Nobecourt,  348 
Nocht,  295 
Noe,  153 
Noeggerath,     23,     335, 

337,  338,  382 
Noguchi,  56 
Nollau,  89 
v.  Noorden,  387 
Norris,  63 
Northrop,  83,  84 
Nutall,  78 

O 

Ochsner,  260  (#773) 

Odake,  171,  172 

Ogata,  291,  329 

Ohler,  92,  250 

Okimoto,  115  (#323), 
271  (#836) 

Okuda,  115  (#323),  271 
(#836) 

Ono,  293 

Oppermann,  308 

Orenstein,  296 

Orgler,  317,  31d 

Osborne,  37,  38,  39,  40, 
41,  43,  44,  74,  89,  90, 
112,  113,  114,  115, 116, 
117,  119,  120,  221, 122, 
123,  125, 127, 136,  188, 
190,  195,  210,  218,  221, 
224,  249,  251,  252,  255, 


257,  258,  262,  264,  265, 
266,  267,  268,  269,  270, 
271,  272,  346,  348,  382, 
393,  395 

Oseki,  245,  264,  265,  266 

O'Shea,  305 

Ostheimer,  313 

Otabe,  60 

Ottow,  103,  196,  244 


Paal,  220 

Pacini,  57 

Padua,  393 

Paget,  311 

Pagniez,  293 

Paguchi,  104 

Palmer,  89,  90,  91,  222, 

257,  368 
Paltauf,  374 
Pappenheimer,  321, 327, 

328 
Park,  E.  A.,  314,  323, 

328 

Park,  F.  S.,  376 
Parker,  279,  382 
Parsons,  44,  73,  134, 

255,  263,  265,  267,  271, 

272,  327,  342,  380 
Partsch,  330 
Parturier,  313 
Pasqualis,  22 
Pasteur,  51 
Patel,  331 
Paton,  140, 323, 325 
Patterson,  372 
Paul,  252,  265,  269 
Paus,  314 
Pearce,  386 
Peckam,  387 
P6hu,  337 
Peiser,  318,  380 
Pepper,  332,  386 
Pe>ronet,  246 
Peters,  80 
Petersen,  364 
Peterson,  154 


Petrone,  333 

Pfaundler,  158,  159, 348 

Pfluger,  136 

Phemister,  320,  323 

Pick,  380 

Pickard,  269 

Pickens,  298 

Pieper,  56 

Pierson,  263, 268, 271 

Pilado,  208 

Pinault,  352 

Pincherle,  333 

Pitz,  110,  129,  130,  221, 

249,    253,    257,    262, 

264,  266 
Place,  148 
Plimmer,  144 
Pol,   32,   95,    192,   243, 

245,  265,  266,  396,  398 
Pommer,  316,  324 
Poole,  62 
Poppe,  253 
Portier,    79,    134,    210, 

214,  398 
Potter,  373 

Poynton,  301,  302,  307 
Praussnitz,  342 
Prescott,  241 
Pringle,  389 
Pringsheim,  H.  H.,  52 
Prinzing,  372 
Pritchard,  260,  324 
Pugliese,  245,  397 

Q 
Queal,  398 

R 

Raczynski,  322 
Rae,  298 
Rahe,  389 

Ramoino,  207,  299,  398 
Ramond,  277 
Ramsden,  396,  398 
Ranc,  79 

Randoin,  79,  134,  210 
Ranwez,  237 


486 


AUTHOR   INDEX 


Ray,  242,  266,  267,  268, 

388 

Rayer,  366 
Reach,  376,  386 
Reakes,  147 
v.  Recklinghausen,  315 
Reed,  279 
Reid,  142 
Reimers,  138 
Re*non,  385 
Report     of     American 

Pediatric       Society, 

392  (#974) 
Report   of  Eng.  Med. 

Res.  Comm.,  118, 175, 

237 
Report  of  Food  Comm. 

of  Royal  Society,  298 
Report    of    Health    of 

City  of  Manchester, 

298 
Report  to  Local  Govt. 

Board,  260 
Reprint  U.   S.   Public 

Health  Service,  260 
Retterer,  313 
Rettger;  56 
Reynolds,  379 
Rheindorf,  303 
Rhodehamel,  189 
Rhodes,  267,  269 
Ribbert,  316 
Richardson,  266 
Richter,  304 
Richet,  77  (#200),  384 
Ricklin,  320 
Riddel,  279,  281 
Ridlon,  361,  366 
Rietschel,  335,  336 
Rimbaud,  279 
Risquez,  397 
Rivers,  62 
Roaf,  367 
Robb,  327,  392 
Roberts,  145,  351,  352, 

353,  356,  360 


Robertson,  50,  84,  125, 

228 

Robinson,  H.  C.,  56 
Robinson,  R.  K.,  85 
Robison,  234 
Roger,  279,  283 
Rogers,  220 
Rohl,  126 
Rohmann,   25,   26,    37, 

125,  126,  127,  245 
Rohmer,  333 
Roily,  308 
Roloff,  138 
Rolph,  352 
Rommel,  145 
Rondoni,  130,  132,  133 

251,  390 
R0nne,  382 
Roper,  338 
Rose,  255,  387 
Rose,  255,  343 
tel  Rosario,  294 
Rosemann,  136 
Rosenfeld,  348 
Rosenheim,  68,  71,  222 
Rosenthal,  347 
Rosin,  307 
Rossi,  239 
Rost,  317 
Roth,  220 
Rothberg,  319,  320 
Rous,  389 
Rouse,  245 
Roussel,  351,  352,  366 
Roxas,  291 
Rubner,    52,   245,   255, 

341,342,376 
Rueck,  311 
Ruh,  260  (#773) 
Rumpel,  350,  372,  373, 

375 

Rumpf ,  292 
Rupp,  57 
Rupprecht,  318 
Russel,  D.  W.,  57 
Russel,  H.  L.,  147 


Rustin,  384 
Rutgers,  343 


Saito,  52 
Salant,  386 
Saleeby,172,206,294 
Salge,  336,  339 
Sambon,  351 
Same1  son,  338 
Sandy,  358 
v.Samson- 

Himmelstiern,  30 
Saneyoshi,  264 
Sato,  297,  303 
Satta,  125 

Sauer,  314,  330,  333,  339 
Saxl,  299  (#954) 
Saxon,  389 
Sazerac,  60 
Schabad,  28,  266,  270, 

314,  317,  318,  319,  320, 

324 

Schaefer,  258  (#754a) 
Schaeffer,  78,  395,  396, 

397 

Schattke,  146 
Schaumann,  20,  32,  36, 

51,  53,  54,   109,   127, 

134, 135, 137, 141, 153, 

155, 170,  172, 189, 196, 

198,  207,  208,  256,  270, 

324,  395 
Schelenz,  297 
Scherer,  295    300,  303, 

305,  309,  333 
Scheunert,  146 
Schiff,  372,  373,  376 
Schilling,  277,  279,  351, 

369 

Schippers,  339 
Schittenhelm,  373,  374, 

375,  376,  377,  388 
Schlapfer,  26  (#30) 
Schlee,  314 
Schlecht  373,  374,  375, 

377,388 


AUTHOR   INDEX 


487 


Schloss,  318, 320 
Schmidt,  125 
Schmitter,  370 
Schmorl,  137,  313,  316 
Schneider,  299  (#954) 
Schnyder,  104 
Schodel,  307,  387 
v.  Schonborn,  109,  215 
Schottelius,  77,  78 
Schreiber,  172,  189 
Schreiner,  70 
Schroder,  297 
Schiiffner,  31,  238,  241, 

279,  363 
Schulhof,  311 
Schumm,  308 
Schut,  293 
Schwartz,  E.,  205 
Schwartz,  H.,  313 
Schwarz,  318 
Scipiades,  329 
Scott,  Agnes  C.,  331 
Scott,  E.  L.,  215 
Scott,  L.  C.,  172 
Seaman,  206 
Sedgwick,  338 
Seefelder,  350 
Segawa,  94,  95 
Seidell,  40, 163, 188, 189, 

193,  196,  398 
Sekine,  260,  270,  272 
Sell,  221,  222 
Sharp,  234 
Sharpe,  140,  334 
Shearer,  58 
van  der  Sheer,  369 
Sheppard,  362 
Sherman,  245,  264,  321, 

327,  342,  366,  398 
Sherwood,  55 
Shiga,  32,  153 
Shimamura,  171,  172 
Shipley,  328 
Shizume,  104 
Shorten,  242,  266,  267, 

268 
Sicard,  279,  283 


Siegert,  324 

Siler,  352,  355,  366 

Simon,  370 

Simmonds,  44,  110,  113, 
130,  193, 195,  221,  223, 
239,  249, 251, 253, 257, 
264,  266,  267,  327,  328, 
342,  380 

Simonini,  317 

Simpson,  32,  186,  293 

Simpson,  F.  R.,  334 

Simpson,  K.,  278 

Sinclair,  313,  392 

Singer,  350 

Sion,  360 

Sittler,  302 

Skelton,  243,  257,  258, 
259,  267,  269 

Skinner,  70 

Slonacker,  114 

Slye,  389 

Van  Slyke,  186 

Smith,  Alice,  H.,  233 

Smith,  C.  A.,  343 

Smith,  C.  H.,  279,  281 

Smith,  D.  W.,  260,  270 

Smith,  H.,  379 

Smith,  M.  L.,  133 

Smith,  W.  J.,  309 

Smith,  Theobald,  128 

Snyder,  353 

Socin,  22,  125 

Sommer,  234 

Sorochowetz,  266,  270 

de  Souza,  202,  203,  344 

Sparkes,  353 

Speroni,  398 

Sperry,  56 

Speyer,  300,  303 

Spiegel,  334 

Spillman,  378 

Spitzer,  257 

Sprawson,  279 

Spriggs,  73 

Spruell,  150 

Stammers,  225 

Stannus,  362 


Stanton,  H.  T.,  30,  31, 
93,  94,  95,  172,  196, 
245,  247,  248,  280 

Stanton,  R.  E.,  361,  362 

Stark,  338 

Stead,  69,  150,  151 

Steenbock,  89,  91,  143, 
150, 193, 221, 222,  223, 
224, 239,  249, 254,  257, 
260,  264, 265, 266,  267, 
268,  270,  271,  343,  396 

Stefansson,  262 

Steinitz,  336,  337 

Stephenson,  222,  383 

Stepp,  20,  21,  36,  96, 
115, 117, 125, 126,  229, 
396 

Sternberg,  395 

Stetter,  314 

Stevenson,  301 

Stevenson,  H.  C.,  55, 
189,  198 

Stewart   146 

Stewart,  C.  A.,  114 

Stewart,  C.  E.,  371 

Stheehman,  332,  333, 
334 

Sticker,  372 

Stiles,  395 

Still,  154,  307 

Stille,  342 

Stilling,  137 

Stockholm,  204 

Stocklasa,  245 

Stolte,  337 

Stolzner,  316,  324,  384 

Strauss,  372 

Strong,  280 

Strongman,  312 

Strudwick,  239 

Stuart,  189 

Stuessy,  259 

Sturtevant,  85,  369 

Sudrez,  251 

Sugiura,  44,  125,  190, 
195,  267,  269,  389 


488 


AUTHOR   INDEX 


Sullivan,  192,  229,  254, 
264,  265, 271, 273,  361, 
362,  365 

Supplee,  40,  258 

Sure,  115 

Sutton,  314 

Suzuki,  T.,  293,  317 

Suzuki,  U.,  115,  171, 
172,  220,  264,  271,  272 

Swanson,  383 

Sweet,  389 

Swett,  257,  368 

Swoboda,  271 

Sydenstricker,  365 

Sylvester,  366 

Szalagyi,  251 

Sztark,  382 


Tachau,  346,  376 
Tada,  319,  320 
Takaki,  29 
Takasu,  333 
Talbot,154,317,337 
Tanaka,  175 
Tanner,  366 
Tasawa,  95 
Taussig,  298,  303 
Taylor,  342 
Telfer,  319 
Teruuchi,  32 
Theiler,   102,   137,   141, 

144, 146,  148, 150, 151, 

210,  211 

Thiemich,  333,  334 
Thierfelder,  78 
Thin,  369 
Thjotta,  62 
Thomas,  E.,  380,  384 
Thomas,  K.,  20,  343 
Thompson,  32,  293 
Thompson,   Helen   B., 

125 

Thorns,  348 
Thurlow,  52 
Tisdall,  334 
Tixier,  316 


Tobler,  301,  320 

Todd,  154 

duToit,  151  (#447) 

Tonin,  372 

Torrey,  129 

Towles,  192,271 

Toyama,  103 

Tozer,  128,  136,  239 

Trappe,  136 

Travis,  279 

Tricoire,  379 

Trop.    Dis.    Bull.,   299 

(#941) 

Tschirch,  398 
Tsujimoto,  220 
Tsuli,  122 
Tsuzuki,  33 
Tiichler,  299  (#954) 
Tullio,  362 
Turk,  328 
Turner,  299 
Tuttle,  352 
Turban,  26  (#30) 

U 

Uhlmann.  209 

Underbill,  25,  140,  385 

Unger,  131,  132,  225, 
234,  235,  240,  242,  243, 
258,  260,  267,  268,  269, 
270,  271, 272, 302,  307, 
310,  311,  314,  321,  322, 
326,328 

Urbeanu,  251 

Urizio,  305 

Utheim,  339 

V 

Vacher,  373 
Valagussa,  380 
Vallery-Radot,  293 
Vandervelde,  372 
Vansteenberge,  52 
Vanutelli,  299 
Vedder,  32,  56,  92,  93, 
103, 145, 172,  206,  207, 


212,  213, 234,  268,  277, 
293,  295,  396 

Viljoen,  102,  137,  141, 
144,  146,  148,  150,  151 
(#447),  210,  211 

Viswallingam,  362 

Vitale,  333 

Vlahuta,  52 

Voegtlin,  96,  97,  109, 
119, 135,  137, 189, 191, 
192, 193, 196,  209, 229, 

245,  246,  249,  254,  264, 
265,  271,  272;  353,  361, 
366,  368,  396 

Voit,  341,  342 
Vokurka,  385 
Volkmann,  315  ( #  1056- 

a) 

Vonderweidt,  333 
Vordermann,  29 

W 
Wakeman,   39,   43,   74, 

112,  136,  190 
Walker,  150 
Wallis,  57,  265,  379 
Walshe,  211,  291 
Walton,  82,  381 
Ward,  109 
Waring,  366 
Washburn,  261 
Wason,  383 
Wassermann,  303,  308 
Waterman,  373 
Watson,   23,   225,   323, 

325 

Watson-Wemyss,  395 
Webster,  186 
Wechuizen,  293 
Wegele,  370 
Weigert,  336,  337,  384 
Weill,  78,  92,  104,  135, 

195,  212, 239, 244, 245, 

246,  248,  254,  264,  271, 
272, 291,  301,  338,  396, 
397 

Weiss,  314,  330,  351,  363 


AUTHOR   INDEX 


489 


Wellman,  92,  172,  353 
Wells,  266,  372,  392 
Werner,  371 
Wernich,  29 
Weston,  353,  373 
Wheeler,  127,  265,  271, 

365,372 

Whipple,238,241,386 
White,  189,  193 
Wieland,  109,  313 
van  der  Wielen,  270 
Wilcock,  24 
Wild,  330 
Wilder,  388 
Wildiers,  51,  52 
Willaman,  64,  65 
Willard,  314 
Willcox,  262,  271,  279, 

294,  299 
Willets,  366 
Williams,  R.  J.,  53,  198, 

199,  201,  202 


Williams,  R.R.,  95,  105, 

Y 

172,  188,  193,  194,  206, 

Yamada,  283 

207,  268,  293,  294 

Yano,  283 

Wilson,  361,  365,  367 

Yellowlees,  351 

Wiltshire,  266,  303,  311 

Yoshikawa,  283 

Winfield,  260,  270 

Winton,  245 

Z 

Wintz,  26  (#30) 

Zak,  305,  375 

Wise,  248 

Zeller,  356 

Wolcott,  279 

Zernik,  350 

Wolf,  60 

Zilva,40,43,55,85,  123, 

de  Wolf,  373 

129,  146,  154,  193,  195, 

ITT*    i/y    on^7 

220,  221,  224,  225,  226, 

Worn,  o97 
Wollmann,  77 
Woltmann,  375 

233,  234,  235,  263,  267, 
270,  307,  311,  328,  380, 
392 

Wood,  353,  357,  363,  369 

Zlocisti,  303 

Woodcock,  384 

Zondek,  373,  375 

Woods,  C.  D.,  252 

Zucker,  321 

Woods,  E.,  245 

Zuntz,  347,  349 

Wrampelmeyer,  40 

Zunz,  331 

SUBJECT  INDEX 


Adenine,      attempt     to      synthesize 

vitamin e  B  from,  193 
Adenoids,  possible  dietetic  origin  of, 

386 
Adrenaline,  in  beriberi,  increase  of, 

133 

in  scurvy,  decrease  of,  133 
Adrenals,  in  scurvy,  enlargement  of, 

133 
Adsorption  of  vitamine  B  from  yeast 

with    colloidal    arsenic    sulfide, 

189 
of    vitamine    B    from    yeast   with 

fullers  earth.   189 
of    vitamine    B    from    yeast    with 

mastic,  189 
of    vitamine    B    from    yeast    with 

norit,  62,  63 

Allantoine,  in  rice  polishings,  170 
Aminoids        (casein       preparation), 

growth  substance  in,  63 
Amphibia,  vitamine  requirements  of, 

85 
Anemia,   influence  of  vitamines  on, 

386 
Animal    experiments    under    sterile 

conditions,  76,  77,  78,  79 
Antiberiberi  vitamine  (see  vitamine 

B) 
Antirachitic  vitamine  (see  Vitamine 

A) 
Antiscorbutic  vitamine  (see  Vitamine 

C) 
Arthritis  deformans,   occurrence  of, 

392 

Artificial  diet,  inadequacy  of,  75 
"Aschamine,"  from  yeast,  190 
Ash-free     diet,     experiments     with, 

21,  22 

Atrophy,  symptoms  of,  336,  337,  339 
therapy  of,  337,  338 


"Auximones,"      similarity      of,      to 

vitamines,  67,  68 
Avitaminoses,  general  discussion  and 

classification  of,  15,  16,  17,  18 
Avitaminosis,  in  cattle,  75,  76 
in  horses,  75,  76 

B 

B.  coli,  growth  of,  60 

B.  diphtheriae,  growth  of,  60,  61,  62 

B.  histolyticus,  growth  of,  60 

B.   influenzae    (B.   Pfeiffer),   growth 

of,  60,  61,  62 

B.  pertussis,  growth  of,  62 
B.  proteus,  growth  of,  60 
B.  sporogenes,  growth  of,  60 
B.  welchii,  growth  of,  60 
B.    xylinum,    growth    of,    on    yeast 

extract,  56 
Bacteria,  certain  types  of,  vitamine- 

containing  medium  for,  57 
probable  function  of,  in  intestinal 

canal,  76,  78 

synthesis  of  vitamine  by,  57 
vitamines    in,    demonstration    of, 

57 
Bacterized  peat,  effect  of,  on  growth 

of  nitrifying  bacteria  of  the  soil 

(B.   radicicola  and  Azotobacter 

chroococcum)  70 

effect    on   growth    of    plants  of 

extract  of,  65,  66,  67 
Banana  fly,  growth  of,  under  sterile 

conditions,  83 

Barley,  nutritive  value  of,  248,  249 
Barlow's  disease,  309   (see  Infantile 

scurvy) 
Beriberi,    effect    of    pilocarpine    on, 

206 

effect  of  pituitary  extract  on,  206 
effect    of    purine    derivatives    on, 

206 


491 


492 


SUBJECT  INDEX 


Beriberi,  effect  of  pyrimidine  deriv- 
atives on,  206 
effect  of  quinine  on,  206 
effect  of  strychnine  on,  206 
effect  of  thyroid  extract  on,  206 
first  clear  conception  as  to  cause  of 

development  of,  31 
human,  acute  pernicious  or  cardio- 
vascular form  of,  289 
human,     chemical     pathology    of, 

293 

human,  dry  atrophic  form  of,  287 
human,  general  pathology  of,  291, 

292 

human,  general  therapy  of,  293 
human,  infantile,  290 
human,  infantile,  therapy  of,  294 
human,  mode  of  occurrence  of  and 

diets  leading  to,  280,  281,  282 
human,  mortality  in,  289 
human,   prevalence  of,  278,  079 
human,  relationship  of,  to  rice  con- 
sumption, 29,  30,  31,  32,  33 
human,  relationship  of,  to  scurvy, 

295 

human,   sensory-motor  form  of,  283 
human,    symptomatology    of,  228, 

283 

in  birds,  blood  sugar  in,  109 
in   birds,    changes   in   the   gastro- 
intestinal tract,  107,  108 
in   birds,    chemical   pathology    of, 

109 
in  birds,  glycogen  content  of  liver 

in,   109 
in  birds,  increase  of  adrenaline  in, 

133 

in  cats,  135 
in  chickens,  92,  93,  94 
in  dogs,  137 
in  pigeons,  97,  98 
in   pigeons,    endocrine   glands    in, 

105,   106,  215 

in  pigeons,  glucose  in,  215 
in  pigeons,  glucosuria  in,  215,  217 
in  pigeons,  heart  in,  104 
in  pigeons,  muscles  in,  104 


in  pigeons,  nervous  system  in,  103, 

104 
in  pigeons,   pathological  anatomy 

of,  103 

in  pigeons,  phlorizin  in,  215 
in   pigeons,    time   of   development 

of,  99 
in  pigeons,  types  of,  99,  100,  101, 

102 
mention    of,    in    Neiching    (oldest 

medical  book  2697  B.  C.)  278 
nature  of  substance  curative  for, 

31,  32,  33 
original   conception   of  cause   and 

prevention  of,  30,  31 
starvation,  211 

"Bios,"  characteristics  of,  51,  52 
effect  of,  on  growth  of  yeast,  55 
nature  of,  controversy  as  to,  52 
similarity  of,   to  vitamine  B,  51, 

52 
Bow  legs,  possible  dietetic  origin  of, 

387 

BreadC 'Soldiers'  bread"),  245 
Butter,  centrifugation  of,  38 


Calculi,    formation    of,    influence   of 

vitamines  on,  393 
Cancer,    influence   of   vitamines   on, 

388,  389,  390,  391 
possibility   of   chemical   substance 

as  etiological  factor  in,  388 
Carbohydrate    dystrophy,    atrophic- 

hydremic  form  of,  335 
etiology  of,  336 
hypertonic  form  of,  335 
pure  atrophic  form  of,  335 
occurrence      of,    in  artificially-fed 

babies,  335 

Carcinomas,  supposed  acceleration  of 
growth  of,  by  underfeeding, 
389 

Carrel's  antiseptic  method  of  wound 
treatment,  possible  use  of  vita- 
mines  as  nutrient  in,  82 
tissue  cultures  in  vitro,  81,  82 


SUBJECT  INDEX 


493 


Carrotinoids,    effect    of   absence    of, 

90,  91,  222,  223 

supposed     relationship  with  vita- 
mine  A,  90 
Casein,   in  hunger  edema,  supposed 

curative  action  of,  376,  377 
nutritive  value  of,  43,  44 
nutritive  value  of,  on  autoclaving, 

344 

Cattle,  "bush-sickness"  in,  147 
diseases  in,   theory  of  poverty  of 

soil  as  cause  of,  147 
"impaction  paralysis"  in,  147 
"lamziekte"  in,  146,  147,  148,  149, 

150,  151 

"stallmangel"  in,  146 
"stijfziekte"  in,  146,  147,  148 
Cats,  beriberi  in,  135 

experimental  rickets   in,    inability 

to  produce,  136 
Cereals,  baking  of,  246 
milling  of,  245 

nutritive  value  and  vitamine  con- 
tent of,   under  various  methods 
of  preparation,  244 
Chicken  beriberi,  92 
sarcoma,    effect   of   vitamines    on, 

390 
Chickens,  growth  of,  importance  of 

vitamines  A  and  B  for,  92 
"leg-weakness"  in,  89 
normal    nutrition    of,    87,    88,    89, 

90,  91,  92 
ophthalmia  in,  87 
polyneuritis  in,  87 
"Cholesterol    gland"     (see    "Lipoid 

gland") 

Choline  in  rice-polishings,  170 
Cod  liver  oil,  chemistry  of,  218 
fractionation  of,  219,  220 
reduction  of,  220 
specificity  of,  for  rickets,  328 
"spinacen"  from,  220 
"squalen"  from,  220 
use   of   for   chemical    isolation    of 
vitamine  A,  218 


Colpidium    colpoda,    growth    of,    in- 
fluence of  vitamine  extracts  on, 
80   ' 
Cooking  water,  vitamine  content  of, 

241 
Corn,   as  cause  of    "emmaisadura," 

250 
consumption    of,    in    relation    to 

pellagra,  250,  251 
dried,   absence  of  vitamine  C  in, 

254 
feeding  of,  as  cause  of  scurvy  and 

beriberi,  250,  251 
nutritive  value  of,  250 
vitamine  content  of,  as  affected  by 

milling,  252,  253 
Coxa  vara,  in  scurvy,  387 

D 

Desmotopa,  growth  of,  84 
Diabetes,  influence  of  vitamines  on, 

387 

limitation  of  diet  in,  possible  pro- 
duction of  avitaminosis  by, 
388 

Diet,  artificial,  inadequacy  of,  75 
ash-free,  experiments  with,  21,  22 
influence    of,    on   growth    of   neo- 
plasms, 389 
weekly,    in   Denmark,    during   the 

War,  349 
weekly,    in    Dresden,    during    the 

War,  348 

Dietary  composition  in  Italian  pella- 
gra districts,  364 
constituents,  necessity  for  proper 

relationship  between,  340 
Dogs,  beriberi  in,  137 
osteoporosis   in,    140 
pellagra-like  condition  in,  140 
rickets  in,  137,  138 
scurvy  in,   137 

scurvy  in,  symptoms  of,  138,  139,  140 
Dropsy,  (see  Epidemic  dropsy) 
Drosophila,   growth  of,   on    solution 
of   cane  sugar  and  salts,  83,   84 


494 


SUBJECT  INDEX 


Drosophila,  ampelophila,  growth  of, 

84 
melanogaster,  growth  of,  84 

E 

Egg  production,  factors  influencing, 

91 
"Emmaisadura,"     rotted     corn     as 

cause  of,  250 
Epidemic  dropsy,  characteristics  of, 

288,  (see  373) 
Essential  food  factors,  opposition  to, 

existence  of,  25 
Exophthalmic    goitre,    as    suspected 

avitaminosis,    380 
Exudative  diathesis,  337,  339 


Ferments,        comparison       between 

vitamines  and,  163 
Fish,  growth  of,  85 
Flat  feet,  possible  dietetic  origin  of, 

387 
Flies,  development  of,  part  played  by 

vitamine  B  in,  84 
Fly,  (see  Drosophila,  Musca  domes- 

tica,    Desmotopa,    Sciaria,    Ty- 

roglypha) 

Foodstuffs,  non-protective  and  pro- 
tective against  rickets,  325 
table  showing  vitamine  content  of, 

264 

Fragilitas  ossium,  in  rickets,  313 
Frogs,  growth  of,   influence  of  vita- 
mines  on,  85,  86,  87 
Fruit  juices,  antiscorbutic  properties 

of,       erroneously      ascribed      to 

laxative  action,  129 
Fullers  earth,  use  of,  in  adsorption 

of    vitamine    B    from    autolyzed 

yeast,    188 
Fungi,     vitamine    requirements    of, 

64,  65 

Aspergillus  niger,  64 
Oidium  lactis,  65 
Sclerotinia  cinera  [(Bon)  Schroter], 

64 


G 

Gal— lamziekte  (See  Lamziekte) 
Goats,     vitamine    requirements    of, 

141 

stijfziekte  and  lamziekte  in,  148 
Gonococcus,  growth  of,  59 
Gossypol,"    toxicity    of,    in    pigs, 

145 
Graves    disease    (see    Exophthalmic 

goitre) 
Guinea  pigs,  vitamine  requirements 

of,  127,  128 
use  of,  in  study  of  scurvy,  127 

H 

"Helles   Mark,"    appearance    of,    in 

scurvy,  132,  309 

Hermeralopia,  as  suspected  avi- 
taminosis, 379 

Hemophilic  bacteria,  factors  neces- 
sary for  growth  of,  similarity  to 
those  required  for  growth  of  rats, 
60 

Higher    plants,    growth    of,    extract 
of  bacterized  peat  in,  67,  68,  69 
"Hikan"  (see  Ophthalmia) 
Horses,  edema  in,  146 

"sukumi"    or     "gokosukumi"    in, 

146 
Hunger  edema,    casein  as  supposed 

curative  for,  in  rats,  376,  377 
differentiation    of,    from    beriberi 

and  scurvy,  377 
development  of,  role  of  adrenaline 

in,  377 

etiology  of,  376 
in  rats,  production  of,  376 
metabolism  in,  375 
mode  of  development  of,  374 
pathology  of,   374 
prevalence  of,  372 
relationship  of,  to  epidemic  dropsy, 

373 

symptomatology  of,  373 
therapy  of,  375 


SUBJECT  INDEX 


495 


I 

Insects,  83 

Intestinal  stasis,   as  suspected   avi- 
taminosis  379 

K 

"Kaffir,"      as      protective      against 

scurvy  300 
Kallak,    as    suspected  avitaminosis, 

378 
Keratomalacia  (see  Ophthalmia) 


Lactalbumin,     nutritive     value     of, 

43,  44,  115 

Lactic  acid  bacteria,  growth  of,  in- 
fluence of  autolyzed  yeast  on, 
52,  53, 

growth  of,  inhibition  of,  53 
Lactose,  supposed  antiscorbutic  ac- 
tion, of  129 

Lamziekte,  etiology  of,  150,  151 
in  cattle  and  goats,  146,  147,  148, 

149 

pathology  of,  149 
six  phases     of,   151 
symptoms  of,  149 
vitamine  B  not  a  causative  factor 

in,  150,  151 

Laxatives,  supposed  antiscorbutic  ac- 
tion of,  129 
Lecithin,  action  of,  due  to  vitamine 

impurity,  81 

influence  of,  on  growth,  81 
in  nutrition,  supposed  influence  of, 

24 
supposed    curative    action    of,    on 

beriberi,  52 

"Lecksucht,"  in  reindeer,  142 
Lemna  minor,   growth  of,   influence 
of    crude    nucleic    acid    deriva- 
tives on,  69 
Leprosy,  as  suspected  avitaminosis, 

385 
"Leptom,"    function    of,    in     plant 

nutrition,  65 
Life  without  bacteria,  76,  77,  78,  79 


Lions,  rickets  in,  136 

"Lipoid  gland,"  in  development  of 

avitaminoses,    relation    of,    229, 

230 
Lipoid  group,  classification  of  those 

substances  essential  to  life  in,  21 
Lipoids,   in  the  body,  synthesis  of, 

20 


M 

Maize  (see  Corn) 

Man,  vitamine  requirements  of,  155, 

156,  157 
Maniok,"  consumption  of,  as  cause 

of  beriberi,  278 
Mammals,   importance  of   vitamines 

for,  109 

Marasmus,  occurrence  of,  337 
"Marmite"   (yeast  extract)  154,  294 
Meat,  significance  of,  as  dietary  com- 
ponent, 262  ' 

"Mehlnahrschaden,"    122,    335    (see 
Carbohydrate      dystrophy     and 
Atrophy) 
Meningococcus,        conditions        for 

growth  of,  57,  58,  59 
probability    of    necessity    of    two 

vitamines  for  growth  of,  58 
Metazoa,  growth  of,  81 
Mice,  vitamine  requirements  of,  125, 

126,  127 
Milk,    in   nutrition,    importance    of, 

256 

nutritive  value  of,  261,  262 
vitamine  content  of,  258 
vitamine  content  of,   influence  of 

drying  on,  260 
vitamine  content  of,   influence  of 

evaporation  on,  260 
vitamine  content  of,   influence  of 

heat  on,  259 

vitamine  content  and  composition 
of,  influence  of  vitamine  content 
of  fodder  on,  256,  257,  258 
vitamines    in,    demonstration    of, 
256,  257 


496 


SUBJECT  INDEX 


Milk,  synthetic,  260 
Milk  dystrophy,  337 
Milk  powder,  ability  of  mice  to  thrive 

on,  22 

Moller-Barlow's  disease,  22  (see  In- 
fantile scurvy) 
Monkeys,  beriberi  in,  153 

pellagra-like  disease  in,   155 

routine  feeding  of,  155 

suitability  of,  for  study  of  scurvy, 

153,  154 

vitamine     requirements     of,     153, 

154,  155 

N 

"Neiching"    (oldest    medical    book, 
2697  B.  C.),  mention  of  beriberi 
in,  278 
Neoplasms,   growth  of,   influence  of 

diet  on,  389,  390,  391 
effect  of  pregnancy  on,  389 
Nicotinic  acid,  attempted  synthesis 

of  vitamine  B  from,  193 
in  rice  polishings,  171 
Nucleic    acid    derivatives,    supposed 
influence  on  growth  of  plants,  70, 
71 
Nucleins,  in  the  body,  synthesis  of, 

20 
Nutrition  in  Europe  during  the  war, 

347,  348,  349 
in  man,  340 

on  high  fat  diet,  338,  339,  346,  347 
on  high  protein  diet,  346,  347 
on  high  starch  diet,  346,  347 
on  high  sugar  diet,  346,  347 
Nutritional  requirements,  73 
Bunge's  statement,  22 
Hopkins'  prophetic  view,  24 

O 

Oats,  nutritive  value  of,  249 
"Oocytin,"   growth  substance  from 

ox-blood,  50 
Ophthalmia,  absence  of  in  Germany 

despite  lack  of  vitamine  A,  349 
as  suspected  avitaminosis,  381,  382 


cure  of,  by  yeast,  117 

cure  of,  by  zinc  sulfate  and  boric 
acid   wash,    117 

in  chickens,  87 

in  rats,    117 

on  diet  containing  butter,  117 
"Opsin,"  in  growth  of  microorgan- 
isms, 56 

'Orypan/'  in  rice  polishings,  172 
"Oryzanin,"  in  rice  polishings  173 
Osteogenesis  imperfecta,  relation  to 

congenital  rickets,  313 
Osteomalacia,  etiology  of,  331 

metabolism  in,  331 

pathology  of,  330 

prevalence  of,  329 

symptoms  of,  330 

therapy  .of,  331 
Osteoporosis,  in  dogs,  140 

relation  to  rickets,  313,  315,  329 


Parabiosis,  in  mice,  125,  126 
Paramecium  aurelia,  growth  of,  effect 
of  vitamine  extract  (pancreas)  on, 
79 

Pellagra,  absence  of,  in  Central 
Europe  despite  small  amount  of 
animal  protein  available,  350 

association  of,  with  the  diet,  363, 
365,  366,  367 

chemical  pathology  of,  361 

condition  in  dogs  similar  to,  140 

etiological  factors  in,  368 

etiology  of,  366 

etiology  of,  in  relation  to  corn  con- 
sumption,  250 

experimental,    production    of,    365 

infantile,  368 

in   monkey,    supposed    production 
of,  367 

lack  of  protein  as  cause  of,  unlike- 
lihood of,  347 

metabolism  in,  362 

mode  of  development  of,  363 

occurrence   of,    ratio   in   men   and 
women,  352 


SUBJECT   INDEX 


497 


Pellagra,  prevalence  of,  351,  352 
prognosis,  362 
progress  of,  352,  353 
relationship    of,    to    beriberi    and 

scurvy,  362,  363 
study  of,  importance  of  pre-pella- 

grous  diet  in,  365 
symptomatology  and  pathology  of, 

355 

bones,  360 

circulatory  system,  359,  360 

endocrine  glands,  361 

eyes,  361 

gastro-intestinal  tract,  355 

nervous  system,  357 

4 'pellagra  glove,"  357 

"pellagra  shoe,"  357 

sexual  organs,  360 

skin,  357 
therapy  of,  366 
types  of,  353 

acute,  malignant,  354 

chronic,  354 

light,  sub-chronic,  354 

severe,     cachectic,     sub-chronic, 

354 
Phosphorus  insufficiency,  theory  of, 

as   explanation   of   beriberi   and 

similar  diseases,  20 
"Phyllanthus  emblica,"   as  cure  of 

scurvy,  297 
Phytin,  in  beriberi,  supposed  value 

of,  33 

Pica,  cure  of,  152 
etiology  of,  152 

in  cattle  and  goats,  147,  148,  151 
in  ostriches  and  poultry,  151 
Pigs,  growth  of,  influence  of  alfalfa 

grass  on,  145 

growth  of,  influence  of  corn  on,  143 
growth  of,  influence  of  wheat  on, 

143 
nutritive'requirements  of,  142,  143, 

144,  145 
susceptibility  of,  to  development  of 

experimental  rickets,  146 


vitamine  requirements   of,    uncer- 
tainty as  to,  146 
Pigeons,  beriberi  in,  97,  98 
beriberi  in,  time  of  development  of, 

99 
beriberi  in,  types  of,  99,  100,  101, 

102 
nutritional    requirements    of,    95, 

96,  97 
Plant    extracts,    growth    promoting 

effect  of,  65 

Pneumonia,  influence  of  diet  on,  385 
Poisons,    toxicity    of,    influence    of 

vitamines  on,  386 
"Polyneuritis    gallinarum,"    21,    30, 

87 
substitution  of  by   "experimental 

beriberi  of  animals,  '  92 
Potatoes,  nutritive  value  and  vita- 
mine  content  of,  255,  256 
"Protein-free  milk,"  as  an  unknown 
factor  in  nutrition,  115 
inadequacy  of,  116,  127 
Protein    minimum,  341,  342,  343,  350 
Proteins,  biological  value  of,  343 
of   various    origin,    table   showing 

nutritive  value  of,  264 
vegetable  and  animal,  74 
vegetable  and  animal,  comparison 
between  nutritive  value  of,  343 
vegetable  and  animal,  possible  ex-- 
planation   of  difference   in   food 
value  of,  43 
vitamine   sparing   action    of,   214, 

325 

Protozoa,  growth  of,  79,  80 
Pyorrhoea    alveolaris,    in    scorbutic 

guinea  pigs,  392 
Pyridine    derivatives,     in    vitamine 

fractions,  presence  of,  165,  166 
attempts    to    synthesize    vitamine 
B  from,  193 

R 

Rabbits,  unsuitability  of,  for  study 

of  scurvy,  135 
ophthalmia  in,  135 


498 


SUBJECT  INDEX 


Rats,  ability  of,  to  choose  between 
two  diets  one  that  is  adequate, 
114 

ability  of,  to  resume  growth  after 
period  of  repression,  114 

food  intake  of,  influence  of  vita- 
mine  B  on,  119,  120 

food  intake  of,  necessity  of  measur- 
ing, 113,  114 

growth  of,  effect  of  lack  of  vitamine 
A  on,  118,  119,  120,  121,  122 

growth  of,  effect  of  lack  of  vita- 
mine  B  on,  118,  119,  120,  121 

growth  of,  effect  of  small  quantities 
of  milk  on,  116 

normal  growth  curves  of,  compari- 
son of,  122,  123,  124 

preparation  of  diets  for,  112,  113 

types  of  cages  for,  111 

vitamine  requirements  of,  110,  114 

vitamine  requirements  of,  on  diets 
rich  in  protein,   fat  starch  and 
sugar,  345,  346,  347 
Reindeer,  suspected  avitaminosis  in, 

142 

Rice,  changes  in,  as  result  of  milling, 
248 

polished,  ability  of  pigeons  to  live 
on,  248 

phosphorus  content  as  possible  in- 
dicator of  nutritive  value  of, 
31,  32 

removal  from,  on  polishing,  of  sub- 
stance protective  against  beri- 
beri, 30 

Rice-polishings,  active  phosphorus- 
free  solution  from,  168 

attempts  to  isolate  vitamine  B 
from,  172,  173,  174,  175,  176 

"beriberi  vitamine"  from,  169 

chemical  investigation  of,  167 

fractionation  of  (diagram),  169 

nutritive  value  of,  248 

substances  isolated  from,  analyses 

of,  170,  171 

Rickets,  association  of,  with  lack  of 
vitamine  A,  323, 


association  of,  with  lack  of  phos- 
phorus, 323 

association  of,  with  lack  of  sun- 
light, 323 

association  of,  with  a  particular 
dietary,  28 

chemical  pathology  of,  315,  316 

cod  liver  oil  in,  specificity  of,  328, 
329 

congenital,  313 

Czerny-Keller  diet  in,  320 

diagnosis  of,  315 

dietetic  theories  with  regard  to, 
324 

effects  of,  in  later  life,  314,  315 

etiology  of,  323 

foodstuffs  not  protective  against, 
325 

foodstuffs  protective  against,  325 

in  dogs,  137,  138 

in  dogs,  symptoms  of,  138,  139, 
140 

in  breast-fed  children,  314 

in  lions,  136 

metabolism  in,  317,  318,  319 

minimal  quantitative  factors  in, 
24  . 

occurrence  of,  313 

pathological  anatomy  of,  315,  316 

prevalence  of,  312,  313 

symptomatology  of,  315 

therapy  of,  and  its  therapeutic 
effect  on  metabolism,  320,  321, 
322 

vitamine  etiology  of,  324,  325,  326, 
327,  328 

X-ray  in  study  of  therapy  of, 
322,  323 

S 
Salt  mixture,  of  McCollum,  113 

Osborne-Mendel,  113 
Schlatter's  disease  (late  rickets),  314 
Schwann's  sheath,  in  beriberi,  292 
Sciaria,  vitamines  in  growth  of,  84 
Scoliosis,  possible  dietetic  origin  of, 
381 


SUBJECT  INDEX 


499 


Scurvy,  association  of  definite  mode 
of  nutrition  with  etiology  of, 
24,  27 

experimental,  conditions  for  dem- 
onstration of,  131,  132 

experimental,  decrease  of  ad- 
renaline in,  133 

experimental,  metabolism  in,  133, 
134 

experimental,  symptoms  of,  131, 
132 

in  guinea  pigs,  discovery  of,  27, 
128,  129 

in  guinea  pigs,  identity  of,  with 
scurvy  in  man,  130 

infantile,  27,  301,  302,  306,  307 

infantile,  latent  form  of,  306 

in  man,  diagnosis  of,  "white  line" 
characteristic  in,  307 

in  man,  differential  diagnosis  of, 
by  means  of  administration  of 
vitamine  C,  307 

in  man,  hematology  in,  308 

in  man,    metabolism    experiments 
in,  308 

in  man,  mode  of  development  of, 
300,  301 

in  man,  pathology  of,  308,  309, 
310 

in  man,  prevalence  of,  297,  298,  299, 
300 

in  man,  relationship  between  beri- 
beri and,  295,  296 

in  man,  symptomatology  and 
progress  of,  303,  304,  305 

in  man,  therapy  of,  311 
Secretin,  non-identity  of,  with  vita- 
mine  B,  208 

Sheep,  vitamine  requirements  of, 
141 

"staggers"  in,  141,  142 
Ship    beriberi,    occurrence    of,    294, 

295 

Soil  conditions  and  vitamines   (dia- 
gram), 76 
Spasmophilia  (see  Tetany) 


"Spindling  sprout  disease,"  of  pota- 
toes, 69 
Spirochetes,  testicular  substance  in 

cultivation  of,  56 
Sprue,    as    suspected    avitaminosis, 

371 

pathology  of,  370 
prevalence  of,   369 
symptomatology  of,  369 
therapy  of,  370 

Sterility,  as  influenced  by  diet,  379 
Stijfziekte,  in  cattle  and  goats,  146, 

147,  148 

cure  of,  by  change  of  diet,  148,  149 
Streptococci,    growth    of,    influence 
of  tryptophane  fraction  of  some 
proteins   on,   344 
explanation  of,  344 

Streptococcus  hemolyticus,  con- 
ditions necessary  for  growth  of, 
62,  63 

failure     to     grow     on     beef-heart 
extract    decolorized   with   norit, 
62,  63 
growth  of,  on  beef-heart  extracts, 

62 

Substances  essential  to  life,  failure 
to  recognize  the  existence  of,  25, 
26 

Synthetic  diets,  deficiency  of,  ex- 
planation of,  23 

T 

Teeth,  development  of,  influence  of 

vitamines  on,  391,  392 
Testicular  substance,  in  cultivation 

of  spirochetes,  56 
Tetany,  nature  of,  332 
etiology  of,  332 
symptoms  of,  332,  333 
therapy  of,  334 
Thyroid,   influence   of  food  poor  in 

vitamine  on,  105 
Tike-tiki    (rice   polishings    extract), 

294 

Trench  sickness,  as  suspected  avita- 
minosis, 378 


500 


SUBJECT  INDEX 


Tubercle  bacilli,  vitamines  in  growth 

of,   60 
Tuberculosis,  as  influenced  by  diet, 

384 
Tumors,    growth    of,    inhibition    by 

underfeeding,  389 
Tyroglypha,  vitamines  in  growth  of, 

84 


Vegetable     soups,      as     therapeutic 
measure    in    nutritive    disturb- 
ances of  children,  337,  338 
Vibrion  septicus,  growth  of,  60 
Vitamine,  antipellagra,   possible  oc- 
currence   in    animal    protein    of, 
45 

choice  of  the  name,  18,  36 

cycle,  theory  of,  71,  72 

content,     of     bacterial     intestinal 
flora,  210 
cooking  water,  241 

most    frequently    used    foodstuffs, 
table  showing,  264 

various  food-stuffs,     influence     of 
ageing    on,    243 
baking  on,  246 
canning  on,  242 
cooking  on,  238 
drying  on,  241 
heating  on,  238 
storing  on,  243 

natural    difficulties     involved     in 
determination  of,  236,  237 

relative  effect  of  temperature  and 
time  of  heating  in  presence 
and  absence  of  air  on,  239, 
240 

hypothesis  of  Portier,  79 

-like  action  of  certain  substances, 
206 

preparations    on    the   market,    in- 
efficiency of,  393,  394 

requirements,  effect  of  dietary  com- 
position on,  210 

dependence  of,  on  amount  of  food 
assimilated,  213 


-sparing  action  of  protein,  214,  262, 

263,  346 

synthesis  of,  by  bacteria,  57 
theory,    importance    of,    effect    of 

clinical  data  on,  26 
types,  35,  36 

Vitamines,  classification  of,  36 
comparison  of,  with  ferments,  163 
conception  of,  research  leading  to, 

19,  20 
demonstration   of  existence   of  A, 

B  and  C,  40,  41 
definition  of,  by  Willaman,  65 
gradual  recognition  of  existence  of, 

37,  38,  39 

inability  of  animal  body  to  synthe- 
size, 49 
influence  of,  on  anemia,  386 

calculi,  formation  of,  393 

cancer,  388,  389,  390 

carcinomas,  389 

chicken  sarcoma,  390 

development  of  teeth,  391,  392 

diabetes,  387 

neoplasms,  388,  389,  390 

toxicity  of  poisons,  386 
in  animal  kingdom,  role  of,  73 
in  bacteria,  demonstration  of,  57 
in   grain   kernels,    localization    of, 

245,  246 
in   higher  plants,    localization   of, 

49 
in  infections,  significance  of,  380, 

381 
in   nutrition,    relative    importance 

of,  73 

in  pure  state,  possible  action  of,  394 
in  tissues,  210 
in  typhoid  bacilli,  57 
in  vegetable  kingdom,  role  of,  49, 

50,  51 

nomenclature,  39 
practical  aspect  of,  393,  394 
relation  of,  to  condition  of  soil,  75 
synthesis  of,  by  some  plants,  49, 

50 
synthesis  of,  by  yeast,  51 


SUBJECT  INDEX 


501 


Vitamine  A,  chemistry  of,  218 

demonstration    of,    use    of    animal 

tests  in,  225 
effect  of  heat  on,  224 
effect    of    lack    of,    on    growth    of 

rats,  118,  119 

effect  of,  oxidation  on,  224 
effect  of,  reduction  on,  224 
effect  of,  ultra-violet  light  on,  224 
in  butter,  220 

in  cod  liver  oil,  218,  219,  220 
in  fat-free  milk,  223 
in  grasses,  221 
in  green  leaves,  probable  formation 

of,  72,  226 

in  physiology  and  pathology,  sig- 
nificance of,  223 
in  synthesis  of  neutral  fats,  role  of, 

226,  227 

in  vegetables,  221 
nature  of,  222 
possible    differentiation    of,    from 

antirachitic  vitamine,  42 
properties    of,    223 
relation    of,    to    carrotinoids,    222, 

223 
relation  of,  to  edema,  prophylaxis 

of,  227 

relation  of,  to  lipoids,  228,  229 
relation  of,  to  ophthalmia,  229 
relation  of,  to  urinary  calculi, 

227 

solubility  of,  in  water,  223 
stability  of,  224 
Vitamine  B,  adsorption  of,  from  auto- 

lyzed  yeast,  with    colloidal    ar- 
senic sulfide,  189 
with  fuller's  earth,   188 
with  mastic,  189 
adsorptive  capacity  of  proteins  for, 

43 
attempted  isolation  of,   from  rice 

polishings,  172, 173, 174, 175, 176 
attempted  isolation  of,  from  yeast, 

177 


attempted  synthesis  of,  from 
adenine,  193 

attempted  synthesis  of,  from  nico- 
tinic  acid,  193 

attempted  synthesis  of,  from  o- 
oxypyridine,  193 

attempted  synthesis  of,  from  p- 
oxynicotine,  193 

as  possible  mother-substance  of 
product  of  endocrine  glands,  208 

chemistry,  physiology  and  pharma- 
cology of,  163,  207 

demonstration  and  estimation  of, 

195 

animal  tests,  195,  197,  198 
phenol  reaction,  197 
phosphorus  content,   196 
nitrogen  content,  196 
uric  acid  reaction,  197 
yeast  fermentation  test,  198 
yeast  growth  test,  198,  199,  200, 
201 

suitability  of  for  water-soluble 
vitamines  but  not  for  vita- 
mine  B,  203 

effect  of,  on  scurvy,  41 

effect  of  lack  of,  on  growth  of  rats, 
118,  119 

fractionation  of,  difficulties  in- 
volved in,  164,  165 

in  carbohydrate  metabolism,  role 
of,  212 

influence  of,  on  food  intake  of  rat, 
119 

in  yeast,  theory  of  occurrence  of, 
as  nucleoproteid,  189,  190 

its  possible  differentiation  from  the 
growth-promoting  substance,  41, 
42,  44,  53,  203,  204 

nature  of,  means  for  studying, 
165,  166,  167 

non-identity  of,  with  secretin,  209 

preparation  of,  from  yeast,  as  ac- 
tive crystalline  substance  of  con- 
stant melting  point,  165  178,  179, 
180,  181 
analyses  of,  182,  183,  184 


502 


SUBJECT  INDEX 


Vitamine  B,  animal  tests  with,  185 
solubility  of,  194 

in  olive  oil,  191 
sources  of  supply  of,  192 
stability  of,  167 
stability    of,     against    heat,     and 

chemical    and    physical    agents, 

194,  195 
Vitamine  C,  chemistry  and  nature  of, 

231 

effect  of,  on  beriberi,  41 
effect  of,  on  growth,  43 
formation  of,  on  germination  of 

grains,  41,  49,  246 
in  differential  diagnosis  of  scurvy, 

307 
influence  of,  on  growth  and  weight 

of  infants,  158,  159 
stability  of,  235 
various  means  of  fractionation  and 

concentration  of,   232,  233,  234, 

235 
Vitamine   D,    in   growth    of   certain 

bacteria,  205 
in  growth  of  rats,  importance  of, 

205 
in   yeast,    demonstration   of,   204, 

205 
separation    of,    from   vitamine   B, 

204,  205 

W 

War  edema,  unlikelihood  of  lack  of 
protein  as  cause  of,  347 

Wheat,  vitamines  in,  localization  of, 
249 


X-ray,  in  diagnosis  of  scurvy,  307 
in  study  of  therapy  of  rickets,  322, 
323 


Yeast,    active   crystalline   substance 

obtained  from,  178,  179,  180,  181 
analysis  of,  182,  183,  184    . 
animal  tests  with,  185 


as  rich  source  of  vitamine  B,  51 

attempts  to  isolate  vitamine  B 
from,  177 

chemical  investigation  of,  177 

curative  influence  of,  on  beriberi, 
51 

dialysis  of,  190 

fermentation  of,  as  affected  by- 
certain  organic  substances,  51, 
52,  53 

growth  of,  influence  of  autolyzed 
yeast  on,  52,  53 

growth  of,  influence  of  size  of 
inoculation,  51 

growth  of,  inhibition  of,  53 

growth  of,  in  presence  of  as- 
paragine,  52 

growth  of,  in  presence  of  peptone, 
52 

growth  of,  in  presence  of  vitamine 
extracts,  52,  54 

growth  of,  method  of  Bachmann, 
54,  198 

growth  of  method,  of  Eddy,  198 

growth  of,  method  of  Funk-Dubin, 
200,  201,  202 

growth  of,  method  of  R.  J.  Wil- 
liams, 53,  54,  199 

growth  of,  not  a  measure  of  vitamine 
B,  53,  203 

growth  of,  unaffected  by  vita- 
mine  C,  54 

in  growth  of  bacteria,  influence  ofr 
56 

in  nutrition  of  flies,  role  of,  84 

methods  for  fractionation  of,  186, 

187,  188,  189,  190,  191 
substance  isolated  from,  184, 185 
substances   isolated    from,    animal 

tests  with,  185 

synthesis  of  vitamine  by,  51,  55 
treatment  of,  with  colloidal  arsenic 

sulfide,  189 
treatment   of,  with   fuller's  earth, 

188,  189 

treatment  of,  with  mastic,  189 
treatment  of,  with  norit,  62,  63 


